RU2340616C2 - Tocopherol-modified therapeutical drug compounds - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: in novel tocopherol-modified therapeutic drug compounds of formula 1 T-L-D, T is tocopherol, L is succinate, and D is camptotecin or its derivative, where all three fragments are bound covalently. Invention also relates to emulsions based on said compounds, formulations of micelles, including said compounds, methods of treating cell proliferative disease using said compounds and formulations, as well as to said compounds application for production of medication for treatment of cell proliferative disease.

EFFECT: increase of composition and treatment method efficiency.

18 cl, 17 dwg, 4 tbl, 19 ex

Description

FIELD OF THE INVENTION

The present invention relates to new therapeutic drugs; compositions that include new therapeutic drugs; and methods of introducing and using new therapeutic drugs and compositions.

BACKGROUND OF THE INVENTION

The ability to administer biologically effective drugs that are poorly soluble in biocompatible solvents to mammals is a major obstacle in the field of pharmaceutical and medical chemistry. In particular, difficulties arise when the active drug is either insoluble in water or unstable in other biocompatible solvents.

A way to increase the solubility of medical agents is to chemically modify them or combine them with another molecule to change the solubility profile in a particular solvent. Associated active drugs, often called prodrugs, include chemical derivatives of biologically active starting materials that are converted to the starting compounds in vivo. The release of the active parent drug from the prodrug may occur as a result of processes such as hydrolysis or enzymatic hydrolysis. The release rate depends on several factors, including the type of chemical bond connecting the active parent drug to the conjugate fragment.

The incorporation of a water-soluble fragment (e.g., polyethylene glycol, polyglutamate, or polymer) to increase the solubility and circulation time of the drug has been investigated by others. The use of fatty acids to connect with active drugs to determine tumors has also been investigated as a way to improve the therapeutic index.

Many potent drugs, such as camptothecin and its analogues (e.g. 10-hydroxycamptothecin and 7-ethyl-10-hydroxycamptothecin), taxanes (e.g. paclitaxel, dodetaxel), candesartan, amphotericin B, azathioprine, cyclosporine, entacapone, danazole, Bosentan, to name a few, is poorly soluble or has poor cellular permeability. Solubility problems of potential therapeutic agents are common and often cause delays in drug development. Several technologies have been developed to facilitate the delivery of poorly soluble and insoluble compounds to patients. Examples of technologies specifically designed to solve solubility problems include complexing agents, nanoparticles, microemulsions, solubility enhancing formulations, prodrugs, and new polymer systems.

Paclitaxel (see structure below), a natural product found in the bast of the Pacific yew, is an example of an important chemotherapeutic agent with a wide spectrum of activity against solid tumors, primary breast cancer, ovarian cancer, colon and large cell lung cancer.

Paclitaxel exerts antitumor activity by binding to tubulin and stabilizing microtubules and thus blocking cell mitosis. However, paclitaxel, like many other potent biologically active molecules, has very limited aqueous solubility.

Camptothecin (CBT) (see structure below) is another example of a poorly soluble and difficult-to-formulate anti-cancer drug.

CBT is a quinoline-based alkaloid found in the cortex of Chinese camptotheca and Asian nothapodytes. The CBT includes four flat rings (ABCD) and one ring in the boat conformation (E). CPT has been found to have a wide spectrum of antitumor activity, especially in solid human tumors. However, the lactone (ring E) of camptothecin and its derivatives is completely mobile under alkaline conditions and physiological pH. Opening of this ring with the formation of particles of an acid salt or carboxylate leads to a significant loss of anticancer activity. Since the early 1960s, when Wall and Wani discovered CBT, efforts have been made to improve the anticancer activity of camptothecin and its analogs and reduce undesirable toxicity. To date, no successful camptothecin formulation has been developed due to its poor solubility in water and organic solvents. However, the water-soluble analogues of camptothecin, irinotecan hydrochloride (CAMPTOSAR) and topotecan hydrochloride (HYCAMPTIN) have been developed and are the only camptothecin analogs currently approved by the Food and Drug Administration.

Recently, a technology has been developed for the formulation of an emulsion of vitamin E based on α-tocopherol for drug delivery paclitaxel. In this formulation, paclitaxel is dissolved in α-tocopherol and is obtained as an oil-in-water emulsion. However, although paclitaxel is soluble in α-tocopherol, the solubility of other active fragments (including camptothecin and other taxanes) in α-tocopherol is limited. Therefore, there remains a need for new methods that are safe and effective, dissolution and delivery of molecules of a poorly soluble active drug.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides therapeutic drug compounds that have been modified to increase their lipophilicity. Compounds of the invention include one or more therapeutic drug moieties and one or more lipophilic moieties. The therapeutic drug moiety is covalently linked to the lipophilic moiety directly or via a linker moiety. Methods of making modified therapeutic drugs are also provided.

In another aspect of the invention, compositions are provided that include compounds of the invention. In one embodiment, these compositions comprise a compound of the invention, optionally one or more other therapeutic agents, and a lipophilic medium. Methods of making compositions are also provided.

In an additional aspect, the invention provides emulsion and micelle formulations that include a compound of the invention. The formulation of the emulsion includes an oil phase and an aqueous phase. The oil phase includes a compound of the invention and a lipophilic medium. The emulsion may be an oil-in-water emulsion or a water-in-oil emulsion. The micelle formulation includes a compound of the invention and an aqueous phase. Methods for making emulsion and micelle formulations are also provided.

Other aspects also provide methods for administering the compounds of the invention to a subject in need thereof and methods for creating conditions suitable for treatment by administering a compound of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

The above aspects and many of the attendant advantages of the present invention will be easier to appreciate and better understood with reference to the following detailed description, taken in combination with the accompanying drawings, where:

FIGURE 1 schematically illustrates the reaction of d-α-tocopherol and a therapeutic drug containing a carbonyl group to produce a tocopherol-modified therapeutic drug compound.

FIGURE 2 schematically illustrates the functionalization of tocopherol with a chlorocarbonyl group (-C (= O) Cl) and a chlorophosphoryl group (-P (= O) OR ₁ Cl) and the reaction of the obtained acid chloride and the corresponding functionalized therapeutic drug compound to produce a tocopherol-modified therapeutic drug compound .

FIGURE 3 schematically illustrates the functionalization of tocopherol with dicarboxylic anhydride (succinic anhydride) and the reaction of the resulting carboxylic acid and the corresponding functionalized therapeutic drug compound to produce a tocopherol-modified therapeutic drug compound.

FIGURE 4 schematically illustrates the preparation of camptothecin tocopherol succinate.

FIGURE 5 schematically illustrates the preparation of tocopherol succinate 10-hydroxycamptothecin and tocopherol succinate 7-ethyl-10-hydroxycamptothecin.

FIGURE 6 schematically illustrates the preparation of 10,20-di (tocopherol succinate) 7-ethyl-10-hydroxycamptothecin.

FIGURE 7 schematically illustrates the preparation of camptothecin tocopherol succinate containing a polyethylene oxide group.

FIGURE 8 schematically illustrates the preparation of paclitaxel tocopherol succinate.

FIGURE 9 schematically illustrates the preparation of docetaxel tocopherol succinate.

FIGURE 10 schematically illustrates the preparation of camptothecin tocopherol terephthalate.

FIGURE 11 schematically illustrates the preparation of tocopherol cyclohexane-1,2-dicarboxylate 7-ethyl-10-hydroxycamptothecin.

FIGURE 12 is a graph comparing in vivo stability of the lactone form of camptothecin and two typical tocopherol-modified therapeutic drug compounds of the invention (SN2300, camptothecin tocopherol succinate and SN2310, 7-ethyl-10-hydroxycamptothecin tocopherol succinate).

FIGURE 13 is a graph comparing the GI ₅₀ values provided by the NCI for camptothecin, irinotecan hydrochloride (irinotecan) and topotecan hydrochloride (topotecan) with the GI ₅₀ values obtained for two typical tocopherol-modified therapeutic drug compounds of the invention (SN2300 and SN2310) cell lines: H460, HCT-116, HT29 and OVCAR-3.

FIGURES 14A and 14B are graphs of concentration-time values after intravenous injection of 13.8 mg / kg of two typical tocopherol-modified therapeutic drug compounds of the invention (FIGURE 14A, SN2300; and FIGURE 14B, SN2310).

FIGURES 15A and 15B are graphs illustrating tumor growth (mm ³ ) over time in x-ray diffractors treated with saline, irinotecan and two typical tocopherol-modified therapeutic drug compounds of the invention (SN2300 and SN2310) for two different tumor models (FIGURE 15A, NCI-H460 and FIGURE 15B, NT-29).

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

In one aspect, the present invention provides therapeutic drug compounds that are modified to increase their lipophilicity. The compounds of the invention are modified therapeutic drugs. Compounds of the invention include a therapeutic drug moiety and a lipophilic moiety.

In some embodiments, the compounds of the invention comprise more than one therapeutic drug moiety. In some embodiments, the compounds of the invention comprise more than one lipophilic moiety. In other embodiments, the compounds of the invention comprise more than one therapeutic drug moiety and more than one lipophilic moiety.

In some embodiments, a therapeutic drug moiety is covalently linked to a lipophilic moiety through a linker moiety. In other embodiments, a therapeutic drug moiety is directly covalently linked to a lipophilic moiety without a binding moiety.

In one embodiment, the lipophilic moiety is a tocopherol moiety, and the compound is a tocopherol-modified therapeutic drug compound. A tocopherol-modified therapeutic drug compound (or “tocopherol” therapeutic drug compound) has one or more tocopherol moieties covalently linked to the therapeutic drug moiety, or a tocopherol moiety covalently linked to one or more therapeutic drug moieties. As indicated above, the tocopherol moiety is covalently linked to the therapeutic drug moiety directly or via a linker moiety.

In one embodiment, the tocopherol-modified therapeutic drug compounds of the invention can be represented by the general formula (1):

(T-L) _n (T) _m D (1)

in which T represents a fragment of tocopherol (i.e. a typical lipophilic fragment); L is a binder moiety; n is 0, 1, 2 or 3; m is 0, 1, 2 or 3; n + m is 1, 2 or 3; and D is a therapeutic drug moiety. In said embodiment, the compound comprises n TL fragments (i.e., tocopherol-binding fragments) and m tocopherol fragments with the proviso that n + m is 1, 2 or 3. Each TL fragment comprises a tocopherol fragment covalently linked to a therapeutic drug fragment through a connecting fragment. Each of the n T-L fragments is covalently linked to a therapeutic drug fragment through a binding fragment of the T-L fragment. In this embodiment, each of the m tocopherol fragments is directly linked to a therapeutic drug fragment without a binding fragment.

Typical compounds having the formula (1) include compounds in which n is 0 and m is 1, 2 or 3. These compounds have the general formula:

T _m D (2)

In said embodiment, one, two, or three tocopherol fragments are directly covalently linked to a therapeutic drug moiety.

Typical compounds having the formula (1) include compounds in which m is 0 and n is 1, 2 or 3. These compounds have the general formula:

(T-L) _n D (3)

In said embodiment, one, two, or three T-L tocopherol fragments are covalently linked to a therapeutic drug fragment through a binding fragment of the T-L fragment.

Typical compounds having the formula (1) include compounds in which m is 1 or 2 and n is 1 or 2. These compounds have the general formulas:

(T-L) (T) D (4)

(T-L) (T) ₂ D (5)

(T-L) ₂ (T) D (6)

In these embodiments, the compounds of the invention have tocopherol fragments that are directly covalently linked to a therapeutic drug moiety without a binder, and tocopherol fragments that are directly covalently linked to a therapeutic drug moiety through a binder (i.e., T-L fragments).

The compounds of the invention described above include one therapeutic drug moiety and one or more lipophilic moieties (e.g., tocopherol moieties). In other embodiments, the compounds of the invention comprise more than one therapeutic drug moiety. In one embodiment, the compounds of the invention comprise two therapeutic drug fragments. In another embodiment, these compounds include three therapeutic drug moieties. For compounds that include more than one therapeutic drug moiety, the therapeutic drug moieties may be the same or different.

For compounds that include more than one therapeutic drug moiety, therapeutic drug moieties can be incorporated into the compound by any suitable method. In some embodiments, the therapeutic drug moieties can be directly covalently paired (for example, the compound includes a —D — D— or —D — D moiety). In other embodiments, therapeutic drug fragments are separated in the compound by a binder fragment (for example, the compound includes a -DLD or -DLD fragment), a lipophilic fragment (for example, the compound includes a -D-T-D- or -D-T-D fragment) or lipophilic binding fragment (for example, the compound includes a -D-T-LD-, -DTLD or -DL-T-D fragment; or -DLTLD- or -DL-T-LD fragment).

Typical compounds comprising two or three therapeutic drug moieties have the general formula:

(T-L) (D) _p  (7)

in which p is 2 or 3. In said embodiment, two or three therapeutic drug moieties are covalently linked to a linker moiety. In this case, the binder includes many sites for the attachment of a therapeutic drug compound (or a modified therapeutic drug compound). As is clear from formula (7), the linker moiety is also covalently linked to the lipophilic moiety (e.g., a tocopherol moiety). As noted above, the compounds of the invention comprising more than one therapeutic drug moiety may have formulas other than those shown above in formula (7). For example, such a compound may include more than one (e.g., two or three) lipophilic moieties (e.g., tocopherol).

Compounds of the invention include one or more lipophilic moieties and one or more therapeutic drug fragments that are either directly covalently linked or covalently linked through linking fragments.

As used herein, the term “lipophilic moiety” means a chemical moiety having lipophilic or hydrophobic characteristics and which increases the solubility of a therapeutic drug compound in a lipophilic solvent or environment when it is covalently coupled to a therapeutic drug compound to form a compound of the invention. Typical lipophilic moieties suitable for the manufacture of compounds of the invention are described below.

As used herein, the term “therapeutic drug moiety” means a chemical moiety derived from a therapeutic drug compound. Typical therapeutic drug compounds suitable for the manufacture of the compounds of the invention are described below.

As used herein, the term “linking moiety” means an atom or group of atoms that covalently binds, for example, a lipophilic moiety to a therapeutic drug moiety. Typical binders suitable for the manufacture of the compounds of the invention are described below.

Lipophilic modification of therapeutic drug compounds . A therapeutic drug compound may have one or more suitable functional groups or may be modified to include one or more suitable functional groups for covalent binding to a lipophilic moiety. Suitable functional groups include, for example, the following groups: hydroxyl group (—OH), amino group (primary amino group —NH _2, or secondary amino group —NHR ₁ , where R _{1 is} independently selected from H, C _1-6 n-alkyl, C _{3- 12} branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl or aralkyl), thiol group (-SH), carboxyl group (-COOH), aldehyde group (-CHO), isocyanate group (-N = C = O), sulfo (-SO ₃ H), sulfate group (-OSO ₃ H), phosphoric acid group (-ORO ₃ H), phosphonic acid group (- ₃ H ₂ O), galogenallilnaya group, a halogenobenzyl group, a halogenobenzyl group, and substituted epoksiranilgruppa (-CH (O) CH _2).

A therapeutic drug compound can be directly connected to a lipophilic moiety (e.g., a tocopherol moiety) via an ester group (-C (= O) O-), a carbamate group (-OC (= O) NH-), a sulfonate group (-SO ₃ - ), a sulfate group (-OSO ₃ -), a phosphate group (-OPO ₃ R ₁ -, where R _{1 are} independently selected from H, C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _{3- 6} cycloalkyl, substituted or unsubstituted aryl or aralkyl), a phosphonate group (-PO ₃ R ₁ -, where R _{1 are} independently selected from H, C _1-6 n-alkyl, C _3-12 branched o alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl or aralkyl) or an ester group (—O—).

The tocopherol compound, a typical lipophilic compound suitable for the manufacture of compounds of the invention, includes a hydroxyl group (—OH). After modification, the tocopherol compound may be covalently linked to a binder compound that includes one or more reactive functional groups. Suitable reactive functional groups include the following groups: hydroxyl group (—OH), amino group (primary amino group —NH _2, or secondary amino group —NHR ₁ , where R _{1 is} independently selected from H, C _1-6 n-alkyl, C _{3- 12} branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl or aralkyl), thiol group (-SH), carboxyl group (-C (= O) OH), aldehyde group (-CHO), isocyanate group ( -N = C = O), sulfo (-SO ₃ H), sulfate group (-OSO ₃ H), phosphoric acid group (-ORO ₃ H), fosfonovoki lai group (-PO ₃ H ₂₎ galogenallilnaya group, a halogenobenzyl group substituted halogenobenzyl group and an epoxy group (-CH (O) CH _2).

Connecting fragments . As noted above, in some embodiments, the compounds of the invention comprise a lipophilic moiety (eg, a tocopherol moiety) covalently linked to a therapeutic drug moiety through a linker moiety. In addition to the embodiments described above, the tocopherol-modified therapeutic drug compounds of the invention can be represented by the general formula (8):

T-A-R-A'-D (8)

where T is a tocopherol fragment (i.e., a typical lipophilic fragment), D is a therapeutic drug fragment, and A-R-A 'is a binding fragment. It will be appreciated that for formulas (1) and (3) to (7) above, each of which includes a linking moiety L, said linking moiety L in these compounds may be a linking moiety A-R-A '.

In formula 8, groups A and A ′ are independently selected from O, S, SO, SO ₂ , NR ₁ , a carboxylate group (—C (= O) O—), an amide group (—C (= O) NR ₁ -), anhydride group (-C (= O) OC (= O) -), carbamate group (-OC (= O) NH-), carbonyldioxy group (-OC (= O) O) -), ureylene group (-NR ₁ C (= O) NR ₂ -), a phosphate group (-OP (= O) (OR ₁ ) O-), a phosphamide group (-OP (= O) (NR ₁ ) O-), a phosphonate group (-OP ( OR ₁ ) O-), phosphonamide group (-OP (= O) NR ₁ -), sulfate group (-OSO ₂ O-), sulfamide group (-SO ₂ NR ₁ -), sulfonate group (-SO ₃ -) , sulfonamide group (-SO ₂ NR ₁ -) and dicarbonyl group -C (= O) R ₃ C (= O) -, wherein R ₃ is absent or represents vuhvalentny alkyl (e.g., - (CH _{2) n -, n = 1-12)} , substituted alkyl, branched alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl. For the above groups, R represents a divalent group selected from the following groups: alkyl; substituted alkyl, branched alkyl; cycloalkyl; substituted cycloalkyl; heteroalkyl; substituted heteroalkyl; aryl; substituted aryl; aralkyl; substituted aralkyl; amino acid; peptide; polypeptide; protein; mono-, di- or polysaccharide; ethylene glycol oligomer, polyethylene glycol; polyalkylene oxide polymers such as polyethylene oxide and polypropylene oxide; and a polyethylene oxide-polypropylene oxide copolymer. For the above groups, R ₁ and R _{2 are} independently selected from Na ⁺ , K ⁺ , H, C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted aralkyl.

As used herein, the term “aryl” means monocyclic or polycyclic aromatic compounds having from 6 to 14 carbon atoms or heteroatoms, and includes carbocyclic aryl groups and heterocyclic aryl groups. Typical aryl groups include phenyl, naphthyl, pyridinyl, pyrimidinyl, thiazolyl, indolyl, imidazolyl, furanyl and the like. The term “aralkyl” means an alkyl group that is substituted with an aryl group.

The following compounds (compounds 9 to 32) are typical examples of compounds having the formula 8.

The linker moiety (L) and the therapeutic drug moiety (D) can be covalently linked via an ester group. In one embodiment, the therapeutic drug moiety comprises a hydroxyl group that is linked to a carboxyl group of the linker moiety. The linker moiety can be coupled to the tocopherol moiety through an ether group (compound 9), an ester group (compound 10), an amino group (compound 11) or an amide group (compound 12).

In another embodiment, the therapeutic drug moiety (D) includes a carboxyl group that is coupled to the hydroxyl group of the linker moiety (L). The linker moiety can be linked by a tocopherol moiety through an ether group (compound 13), an ester group (compound 14), an amino group (compound 15) or an amide group (compound 16).

In the above compounds, the divalent group R is selected from alkyl; substituted alkyl; branched alkyl; cycloalkyl; substituted cycloalkyl; heteroalkyl; substituted heteroalkyl; aryl; substituted aryl; aralkyl; substituted aralkyl; amino acids; peptide; a polypeptide; squirrel; mono-, di- or polysaccharide; ethylene glycol oligomers; polyethylene glycol; polyalkylene oxide polymers such as polyethylene oxide and polypropylene oxide; and a polyethylene oxide-polypropylene oxide copolymer. In the above compounds, the divalent group R _{1 is} selected from H, C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted aralkyl.

Although a preferred embodiment of the invention is illustrated and described, it should be appreciated that various changes may be made therein without departing from the spirit and scope of the invention. The linker moiety (L) and the therapeutic drug moiety (D) can be covalently linked via an amide group. In one embodiment, the therapeutic drug moiety includes an amino group that is coupled to a carboxyl group of the linker moiety. The linker moiety can be coupled to the tocopherol moiety through an ether group (compound 17), an ester group (compound 18), an amino group (compound 19) or an amide group (compound 20).

In the above compounds, the divalent group R is selected from alkyl; substituted alkyl; branched alkyl; cycloalkyl; substituted cycloalkyl; heteroalkyl; substituted heteroalkyl; aryl; substituted aryl; aralkyl; substituted aralkyl; amino acids; peptide; a polypeptide; squirrel; mono-, di- or polysaccharide; ethylene glycol oligomers; polyethylene glycol; polyalkylene oxide polymers such as polyethylene oxide and polypropylene oxide; and a polyethylene oxide-polypropylene oxide copolymer. In the above compounds, R ₁ and R _{2 are} independently selected from H, C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted aralkyl.

The linker moiety (L) and the therapeutic drug moiety (D) can be covalently linked via an ether group (compound 21) or an amino group (compound 22). In one embodiment, the therapeutic drug moiety includes a hydroxyl group, and in another embodiment, the therapeutic drug moiety includes an amino group.

In the above compounds, R _{1 is} selected from H, C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl or aralkyl.

The tocopherol moiety (T) and the therapeutic drug moiety (D) can be covalently linked via a carbonyldioxy group (—OC (= O) O—) (compound 23). In this case, the linker moiety is a carbonyldioxy group and the therapeutic drug moiety includes a hydroxyl group.

The tocopherol fragment (T) and the therapeutic drug fragment (D) can be covalently linked via the anhydride group (—C (= O) OS (= O) -). In one embodiment, the therapeutic drug moiety comprises a carboxyl group that is coupled to a carboxyl group of the linker moiety. The linker moiety can be coupled to the tocopherol moiety through an ether group (compound 24), an ester group (compound 25), an amino group (compound 26) or an amide group (compound 27).

In the above compounds, the divalent group R is selected from alkyl; substituted alkyl; branched alkyl; cycloalkyl; substituted cycloalkyl; heteroalkyl; substituted heteroalkyl; aryl; substituted aryl; aralkyl; substituted aralkyl; amino acids; peptide; a polypeptide; squirrel; mono-, di- or polysaccharide; ethylene glycol oligomers; polyethylene glycol; polyalkylene oxide polymers such as polyethylene oxide and polypropylene oxide; and a polyethylene oxide-polypropylene oxide copolymer. In the above compounds, R _{1 is} selected from H, C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted aralkyl.

The tocopherol fragment (T) and the therapeutic drug fragment (D) can be covalently linked via a phosphate, phosphoramide or thiophosphate group (compound 28).

In the above compounds, X represents O, NR ₂ or S; X ₁ represents O, NR ₃ or S; R _{1 is} selected from Na ⁺ , K ⁺ , H, C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl or aralkyl; and R ₂ and R _{3 are} independently selected from C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl or aralkyl.

The tocopherol moiety (T) and the therapeutic drug moiety (D) can be covalently linked via a sulfate, thiosulfate or sulfonamide group (compound 29).

In the above compounds, X represents O, NR ₁ or S; and R _{1 is} selected from H, C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl or aralkyl.

A tocopherol moiety (T) and a therapeutic drug moiety (D) can be covalently linked via a ureylene group (—NHC (= O) NH—) (compound 30).

In the above compounds, R ₁ and R _{2 are} independently selected from H, C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl or aralkyl.

The tocopherol fragment (T) and the therapeutic drug fragment (D) can be covalently linked via a carbamate group (—NR ₁ C (= O) O— or —OC (= O) NR ₂ -, compounds 31 and 32, respectively).

In the above compounds, R ₁ and R _{2 are} independently selected from H, C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl or aralkyl.

The connecting fragment . Compounds of the invention include one or more lipophilic moieties. The lipophilic moiety or moieties increase the solubility of the compound in a lipophilic solvent or environment. In one embodiment, the lipophilic moiety is a tocopherol moiety.

As used herein, the term “tocopherol moiety” means a chemical moiety that is derived from a family of natural or synthetic compounds also known by their generic names tocol or vitamin E. In addition to tocopherol compounds, tocotrienol compounds are included in this family. These compounds include a chromatan head having phenolic alcohol (C-6) and a wick tail (C-2). These compounds have the following general formula:

Tocopherols constitute a series of related benzpyranols (or methyltokols) in which the C-2 phytyl (sixteen carbon) side chain of C-2 is saturated. Typical tocopherols include α-tocopherol, (d-form, dl-form, l-form), β-tocopherol (d-form, dl-form, l-form), γ-tocopherol (d-form, dl-form, l-form) and δ-tocopherol (d-form, dl-form, l-form). Among tocopherols, α-tocopherol predominates. Tocotrienols are similar in structure tocopherols except that trienols have three double bonds in the C-2 of the wick side chain.

Compounds of tocopherol and tocotrienol suitable for the manufacture of compounds of the invention include the compounds shown below.

Compound R _one R ₂ R ₃ alpha (α) CH ₃ CH ₃ CH ₃ beta (β) CH ₃ H CH ₃ gamma (γ) H CH ₃ CH ₃ delta (δ) H H CH ₃

As used herein, the term "tocopherol" refers to any member of the tocopherol family mentioned above.

Therapeutic drug fragment . Compounds of the invention include one or more therapeutic drug fragments. In fact, any therapeutic drug compound having a suitable functional group or which can be modified to include a suitable functional group can be covalently linked to a lipophilic compound to form a compound of the invention. Typical functional groups include, for example, hydroxyl groups (—OH), amino groups (primary amino groups —NH ₂ and secondary amino groups —NHR), thiol groups (—SH), carboxyl groups (—COOH), aldehyde groups (—CHO), isocyanate groups (-N = C = O), sulfonic acid groups (-SO ₃ H), sulfuric acid groups (-OSO ₃ H), phosphoric acid groups (-OPO ₃ H), phosphonate groups (-PO ₃ OR ₁ R ₂ ), and R ₁ and R _{2 are} independently selected from H, C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl or aralkyl a, haloallyl groups, halogenbenzyl groups, substituted halogenbenzyl groups and epoxy groups (—CH (O) CH ₂ ).

Therapeutic drug compounds suitable for the manufacture of the compounds of the invention should not be substantially water insoluble, although the tocopherol modification of the present invention is particularly well suited for the preparation and delivery of such water insoluble compounds.

In one embodiment, the therapeutic drug moiety is derived from a therapeutic compound that is substantially insoluble in water. In another embodiment, a therapeutic drug moiety is prepared from a therapeutic compound that is substantially insoluble in organic solvents. In another embodiment, a therapeutic drug moiety is prepared from a therapeutic compound that is substantially insoluble in water and essentially insoluble in organic solvents. In one embodiment, the therapeutic drug compound has a solubility in water at room temperature of less than about 1000 μg / ml. In one embodiment, the therapeutic drug compound has a solubility in water at room temperature of less than about 500 μg / ml. In one embodiment, the therapeutic drug compound has a solubility in water at room temperature of less than about 100 μg / ml. In one embodiment, the therapeutic drug compound has a solubility in water at room temperature of less than about 25 μg / ml.

Typical therapeutic drug compounds suitable for the manufacture of the compounds of the invention include anticancer compounds (e.g., paclitaxel and its derivatives, including docetaxel, camptothecin and its derivatives, including 7-ethyl-10-hydroxycamptothecin (SN38) and 10-hydroxycamptothecin, and doxorubicin and its derivatives), antifungal compounds (e.g., flucanazole), antibacterial compounds (e.g., penicillin G, penicillin V), antihypertensive compounds (e.g. hydralazine, candesartan and carvediol), a counterweight inflammatory compounds (e.g., isoxicam), antidiabetic compounds (e.g., metformin), antiviral compounds (e.g., lamivudine), antidepressant compounds (e.g., fluoxetine), antihistamines (e.g., hydroxyzine), antiarrhythmic compounds (e.g., procainamide hydrochloride), antihyperlipope compounds (e.g. probucol) and compounds for reproductive health (e.g. danazol) and the treatment of Parkinson's disease (e.g. lazabemide) and immunosuppressive (e.g. azathioprine and cyclo sporin) and respiratory (e.g., bosentan) diseases and conditions. Other therapeutically useful biological materials that can be modified according to the invention include biologically active proteins, enzymes and peptides.

In one embodiment, the therapeutic drug moiety is derived from an anti-cancer compound. Typical anti-cancer therapeutic compounds include taxanes. Taxanes include any anti-mitotic taxane, taxane derivative, or equivalent. As used herein, the term "taxane" refers to taxanes, taxins, and taxoids in the same way as their derivatives or analogs.

Paclitaxel and its derivatives and analogues are members of the taxane family. Paclitaxel derivatives include, for example, paclitaxel benzoate derivatives such as 2-debenzoyl-2-aroyl and C-2-acetoxy-C-4-paclitaxel, 7-deoxytaxol, C-4 aziridine paclitaxel, as well as various paclitaxel compounds with natural and synthetic polymers, especially with fatty acids, phospholipids and glycerides, and 1,2-diacyloxypropan-3-amine. Other derivatives of paclitaxel include docetaxel; spicatin; taxane-2,13-dione, 5β, 9β, 10β-trihydroxy-, cyclic 9,10-acetal with acetone, acetate; taxane-2,13-dione, 5β, 9β, 10β-trihydroxy-, cyclic 9,10-acetal with acetone; taxan-2β, 5β, 9β, 10β-tetrol, cyclic 9,10-acetal with acetone; taxan; cephalomannin-7-xyloside; 7-epi-10-deacetylcephalomannine; 10-deacetylcephalomannine; cephalomannine; Taxol B; 13- (2 ', 3'-dihydroxy-3'-phenylpropionyl) baccatin III; unnanxol; 7- (4-azidobenzoyl) baccatin III; N-debenzoyltaxol A; O-acetylbaccatin IV; 7- (triethylsilyl) baccatin III; 7,10-di-O - [(2,2,2-trichloroethoxy) carbonyl] baccatin III; baccatin III 13-O-acetate; baccatin diacetate; baccatin; baccatin VII; baccatin VI; baccatin IV; 7-epi-baccatin III; baccatin V; baccatin I; baccatin III; baccatin A; 10-deactyl-7-epitaxol; epitaxol; 10-deacetyltaxol C; 7-xylosyl-10-deacetyltaxol; 10-deacetyltaxol-7-xyloside; 7-epi-10-deacetyltaxol; 10-deacetyltaxol; or 10-deacetyltaxol B.

Other anticancer compounds suitable for the manufacture of the compounds of the invention include camptothecin and its derivatives, including 7-ethyl-10-hydroxycamptothecin (SN38) and 10-hydroxycamptothecin, and doxorubicin and its derivatives.

In some embodiments, the therapeutic drug moiety is derived from paclitaxel, docetaxel, camptothecin, or derivatives thereof.

For compounds of the invention having formula (2) with m = 1, formula (3) with n = 1 and formula 8, certain compounds are excluded and are not included in the scope of the invention. When the binder moiety is 2-hydroxypropylene (—CH ₂ CH (OH) CH ₂ -), the therapeutic drug moiety is not an α-amino acid (e.g., glycine, alanine, proline, cysteine, aminobutyric acid, aspartic acid, glutamic acid), ω-amino acid (e.g., β-alanine, γ-aminobutyric acid, ε-aminocaproic acid, 2-aminoethanesulfonic acid (taurine)) or a peptide containing a cysteine residue linked through its N-terminus or thiol group (e.g., glutathione) . When the linker moiety is succinate, the therapeutic drug moiety is not an S-linked amino compound or amino acid linked to one of the aliphatic carbons of the succinate. When the linker moiety is succinate, the therapeutic drug moiety is not ferulic acid or its ester.

In another aspect, methods for the manufacture of compounds of the invention are provided. There are many ways to covalently bind a lipophilic compound (e.g., a tocopherol compound) with a therapeutic drug compound to form a compound of the invention. In one embodiment, a typical tocopherol, d-α-tocopherol, includes a hydroxyl group that can be directly linked to the carbonyl group of a therapeutic drug to form a tocopherol-modified therapeutic drug compound. The preparation of a typical tocopherol-modified therapeutic compound of the invention from a therapeutic drug compound containing a carboxylic acid is shown in FIGURE 1.

In another embodiment, the tocopherol can be functionalized on the hydroxyl group with a reagent for introducing an active group, such as phosphoric acid chloride (-P (O) OR ₁ Cl), phosphonic acid chloride (-P (O) R ₁ Cl), sulfonyl chloride (-SO ₂ Cl) or carbonyl chloride (-COCl). The resulting acid chloride can then be reacted with an appropriate functionalized therapeutic drug compound to produce a tocopherol-modified therapeutic drug compound.

In FIGURE 2, X is O, S or NH; and R _{1 is} independently selected from H, C _1-6 n-alkyl, C _3-12 branched alkyl, substituted or unsubstituted C _3-6 cycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted aralkyl.

In another embodiment, the tocopherol can be functionalized on the hydroxyl group with a dicarboxylic acid, ester or anhydride. Suitable reagents include succinic acid anhydride, 1,2-cyclohexanedicarboxylic anhydride, 2,3-dimethyl succinic anhydride, 3,3-tetramethylene glutaric anhydride, glutaric anhydride, maleic anhydride, phthalic anhydride, terephthalic acid or isophthalic acid for COOH). The resulting carboxyl group can then directly react with the corresponding functionalized therapeutic drug, or the carboxyl group can be converted to a more reactive carbonyl chloride group (-COCl), and then the carbonyl chloride group can combine with the functional group of the therapeutic drug to form a tocopherol-modified therapeutic drug compounds as shown in FIGURE 3. In FIGURE 3, X denotes O, S, NH or C (= O) O.

In another embodiment, the binder may be coupled to the hydroxyl group of the tocopherol, and then the therapeutic drug may be attached to an available functional group of the binder. The functional group may be, for example, but not limited to, a carboxyl group (—COOH), a polyethylene oxide group (- (CH ₂ CH ₂ O) _n —H), an aldehyde group (—CHO), an isocyanate group (—N = C = O ), a phosphoric acid group (-OPO ₃ H ₂ ) or a phosphoric acid chloride group (-OPO ₂ R ₁ Cl, where R ₁ is substituted or unsubstituted alkyl or cycloalkyl, substituted or unsubstituted aryl or aralkyl), phosphonic acid chloride group (-PO ₂ R ₁ Cl, where R ₁ represents substituted or unsubstituted alkyl or cycloalkyl, substituted or unsubstituted aryl or aralkyl), a sulfuric acid group (—OSO ₃ H ₂ ), a chlorosulfonic group (—SO ₃ Cl), or an epoxy group (—CH (O) CH ₂ ).

The syntheses of typical tocopherol-modified therapeutic drug compounds of the invention are shown in FIGURES 4 to 11 and described in examples 1 to 13.

FIGURE 4 illustrates the preparation of tocopherol succinate camptothecin compound. Tocopherol succinic acid (Vitamin E succinic acid) has a free carboxyl group which can be coupled to a hydroxyl group, an amino group, a thiol group or a carbonyl chloride group to produce a tocopherol-modified therapeutic drug having a succinate group as a binder. In FIGURE 4, the carboxyl group of the tocopherolosuccinic acid is coupled to the hydroxyl group of camptothecin. The preparation of camptothecin tocopherol succinate is described in Example 1.

FIGURE 5 illustrates the preparation of tocopherol succinate 10-hydroxycamptothecin and tocopherol succinate 7-ethyl-10-hydroxycamptothecin (SN38). Tocopherolosuccinic acid is converted to the corresponding chloride and then reacted with 10-hydroxycamptothecin or 7-ethyl-10-hydroxycamptothecin (SN38). The preparation of tocopherol succinate 10-hydroxycamptothecin and tocopherol succinate 7-ethyl-10-hydroxycamptothecin are described in examples 2 and 3, respectively.

FIGURE 6 illustrates the preparation of 10,20-di (tocopherol succinate) SN38, which contains one therapeutic drug (SN38) fragment, two tocopherol fragments and two binding fragments (succinyl groups). The preparation of 10,20-di (tocopherol succinate) SN38 is described in Example 4.

Suitable binder fragments may include an oligomer or polymer, such as a peptide, polypeptide, protein, mono-, di- or polysaccharide, ethylene glycol oligomer, polyethylene glycol, polyalkylene oxide, such as polyethylene oxide and polypropylene oxide; and a polyethylene oxide-polypropylene oxide copolymer.

FIGURE 7 illustrates the preparation of a tocopherol-modified camptothecin containing a binder moiety that includes a polyethylene oxide group. The preparation of camptothecin tocopherol succinate having a binder moiety that includes a polyethylene oxide group is described in Example 5.

FIGURE 8 illustrates the preparation of paclitaxel tocopherol succinate. During this preparation, tocopherol succinic acid is converted to the corresponding acid chloride and then reacts with paclitaxel. The preparation of paclitaxel tocopherol succinate is described in Example 6.

FIGURE 9 illustrates the preparation of docetaxel tocopherol succinate. The preparation of docetaxel tocopherol succinate is described in Example 7.

FIGURE 10 illustrates the preparation of camptothecin tocopherol terephthalate. During this preparation, tocopherol first combines with terephthalate to form tocopherol terephthalate (Example 9) and then combines with camptothecin to form camptothecin tocopherol terephthalate. The preparation of camptothecin tocopherol terephthalate is described in Example 10.

FIGURE 11 illustrates the preparation of tocopherol cyclohexane-1,2-dicarboxylate SN38. The preparation of tocopherol cyclohexane-1,2-dicarboxylate SN38 is described in Example 11.

The preparation of doxorubicin tocopherol succinate and hydroxyzine tocopherol succinate is described in examples 12 and 13, respectively.

In another aspect, the present invention provides compositions that include compounds of the invention. These compositions include one or more compounds of the invention, optionally one or more additional therapeutic agents, and a lipophilic medium. In one embodiment, the tocopherol-modified therapeutic drug compound is dissolved in a lipophilic medium. Due to the lipophilic fragment, this compound has improved lipophilicity compared to the unmodified therapeutic drug compound. The lipophilic medium (or carrier) of the composition can be any medium from a variety of lipophilic media, including, for example, oils. In one embodiment, the lipophilic medium comprises tocopherol (e.g., α-tocopherol). Typical oils suitable as a lipophilic medium include the following:

Fatty acids and their esters, including carboxylic acids with different chain lengths, mainly straight chain, but which can be branched, examples of which include capric, caprylic, caproic, lauric, myristic, stearic, oleic, linoleic, behenic, as are saturated and unsaturated fatty acids and esters.

Fatty acids esterified with glycerol to form mono-, di- or triglycerides, which may be synthetic or derived from natural sources, including but not limited to, for example, glycerides such as soybean oil, cottonseed oil, rapeseed oil, fish oil, castor oil oil, Capmul MCM, Captex 300, Miglyol 812, glyceryl monooleate, triacetin, acetylated monoglyceride, tristearin, glyceryl behenate and diacetyl tartaric acid esters of monoglycerides.

Glycerides associated with other fragments, such as polyethylene glycol (e.g. Labrasol, Labrafac, Cemophor EL).

Phospholipids, natural or synthetic, such as dimyristylphosphatidylcholine, egg lecithin and pegylated phospholipids.

Other fatty esters, including fatty alcohols (myristyl myristate, isopropyl palmiate) or sugars (sorbitol monooleate, SPAN 80, Tween 80, sucrose laurate).

Fatty alcohols, such as stearyl alcohol, lauryl alcohol, benzyl alcohol, or their esters or ethers, such as benzyl benzoate.

Fat-soluble vitamins and derivatives, for example vitamin E (including all tocopherols and tocotrienols and derivatives of tocopherol and tocotrienol, such as vitamin E succinate, vitamin E acetate and vitamin E succinate polyethylene glycol (TPGS)).

Organic co-solvents can also be used in compositions, optionally in combination with water, including, for example, ethanol, polyethylene glycol, propylene glycol, glycerin, N-methylpyrrolidone and dimethyl sulfoxide.

The solubilities of two typical tocopherol-modified camptothecin compounds of the invention in several media are compared with camptothecin in Example 14.

In an additional aspect, the invention provides emulsion, microemulsion and micelle formulations that include a compound of the invention. Methods for preparing emulsion, microemulsion and micelle formulations are also provided.

As used herein, the term “emulsion” refers to a colloidal dispersion of two immiscible liquids, such as oil and water, in the form of droplets, whose diameter is generally between 0.1 and 3.0 microns, and which are usually optically opaque, if only refractions of the dispersed and continuous phases do not coincide. Such systems have limited stability, usually determined by the application or significant reference system, which can be enhanced by the addition of amphiphilic molecules or viscosity enhancers.

The term "microemulsion" refers to a thermodynamically stable, isotropically transparent dispersion of two immiscible liquids, such as oil or water, stabilized by a surface film of surfactant molecules. The microemulsion has a main droplet diameter of less than 200 nm, typically from 10 to 50 nm. In the absence of water, mixtures of oil (villages) and a non-ionic surfactant (s) form transparent and isotropic solutions, which are known as self-emulsifying drug delivery systems (SESDS) and can be used to improve lipophilic drug dissolution and oral absorption.

Emulsion and microemulsion formulations include an oil phase and an aqueous phase. The emulsion or microemulsion may be an oil-in-water emulsion or a water-in-oil emulsion. The oil phase includes one or more compounds of the invention and a lipophilic medium as described above. In one embodiment, the compound is present in the formulation in an amount of from about 0.005 to about 3.0 weight percent of the total weight of the formulation. In one embodiment, the compound is present in the formulation in an amount of from about 0.01 to about 2.5 weight percent of the total weight of the formulation. In one embodiment, the compound is present in the formulation in an amount of from about 0.1 to about 1.5 weight percent of the total weight of the formulation. In one embodiment, the lipophilic medium is present in the formulation in an amount of from about 2 to about 20 weight percent of the total weight of the formulation. In one embodiment, the lipophilic medium is present in the formulation in an amount of from about 4 to about 12 weight percent of the total weight of the formulation. In one embodiment, the lipophilic medium is present in the formulation in an amount of from about 6 to about 10 weight percent of the total weight of the formulation.

In one embodiment of an emulsion or microemulsion, the compound is a tocopherol-modified therapeutic drug compound, the lipophilic medium comprises tocopherol, and the aqueous medium is water.

In addition to the compounds of the invention, emulsion or microemulsion formulations may include other components commonly used in emulsions and microemulsions and especially used in pharmaceutical emulsions and microemulsions. These components include, but are not limited to, surfactants and co-solvents. Typical surfactants include nonionic surfactants, such as tocopherol surfactants and surfactants.

Suitable surfactant derivatives of tocopherol include tocopherolpolyethylene glycol derivatives such as vitamin E polyethylene glycol succinate (e.g. d-α-tocopherolpolyethylene glycol 1000 succinate, TPGS), which is a derivative of vitamin E in which the polyethylene glycol is attached with an acid hydroxy silane E. The term “Vitamin E polyethylene glycol succinate” as used herein includes Vitamin E polyethylene glycol succinate and derivatives of Vitamin E polyethylene glycol succinate, and eyuschie various communication esters and ethers. TPGS is a nonionic surfactant (HLB = 16-18). TPGS has been reported to inhibit P-glycoprotein, a protein that contributes to the development of resistance to many drugs. Embodiments of formulations of the invention that include TPGS therefore include a P-glycoprotein inhibitor. Surfactant tocopherol derivatives (e.g. TPGS) may be present in the formulations of the invention in an amount of from about 1 to about 10 weight percent, from about 2 to about 6 weight percent, or about 5 weight percent of the total weight of the formulation.

Suitable nonionic surfactants include block copolymers of ethylene oxide and propylene oxide, known as POLOXAMERS and PLUROINICS. These synthetic block copolymers have the general structure: H (OCH ₂ CH ₂ ) _a (OC ₃ H ₆ CH ₂ ) _b (OCH ₂ CH ₂ ) _a OH. The following options, based on values a and b, are commercially available from BASF Performance Chemicals (Parsippany, New Jersey) under the PLURONIC trademark and consist of a group of surfactants designated CTFA as POLOXAMER 108, 188, 217, 237, 238, 288, 338 , 407, 101, 105, 122, 123, 124, 181, 182, 183, 184, 212, 231, 282, 331, 401, 402, 185, 215, 234, 235, 284, 333, 334, 335 and 403 For the most commonly used POLOXAMERS 124, 188, 237, 338 and 407, the values of a and b are 12/20, 79/28, 64/37, 141/44 and 101/56, respectively. In one embodiment, nonionic surfactants are present in the formulation in an amount of from about 0.5 to about 5 weight percent of the total weight of the formulation.

Co-solvents suitable for formulations include, but are not limited to, ethanol, polyethylene glycol, propylene glycol, glycerin, N-methylpyrrolidone, dimethylamide, and dimethyl sulfoxide. Polyethylene glycol (PEG) is a hydrophilic polymerized form of ethylene glycol consisting of repeating units having a chemical structure: (-CH ₂ CH ₂ O-). The general formula for polyethylene glycol is H (OCH ₂ CH ₂ ) _n OH. The molecular weight varies from 200 to 10,000. Such various forms are described by their molecular weight, for example, PEG-200, PEG-300, PEG-400 and the like.

Paclitexel emulsions and their components are described in US Pat. No. 6,458,173 and US Pat. No. 6,660,286, each of which is specifically incorporated herein by reference.

Typical emulsions including tocopherol-modified therapeutic drug compounds (for example, docetaxel tocopherol succinate, paclitaxel tocopherol succinate, camptothecin tocopherol succinate, 7-ethyl-10-hydroxycamptothecin-tococycotocin and 10 tocopherocycocotocin are described in Example 15). drug compounds (for example, 7-ethyl-10-hydroxycamptothecin and tocopherol succinate and camptothecin tocopherol succinate) are described in Example 16.

In an additional aspect, the invention provides micelle formulations that include a compound of the invention and an aqueous phase. Micelles are organized aggregates of one or more surfactants in solution. In one embodiment, the compound is present in the formulation in an amount of from about 0.005 to about 3.0 weight percent of the total weight of the formulation. In one embodiment, the compound is present in the formulation in an amount of from about 0.01 to about 2.5 weight percent of the total weight of the formulation. In one embodiment, the compound is present in the formulation in an amount of from about 0.1 to about 1.0 weight percent of the total weight of the formulation. Suitable surfactants include the above surfactants and in amounts specified above. In one embodiment of a micelle formulation, the compound is a tocopherol-modified therapeutic drug compound, and the surfactant is tocopherolpolyethylene glycol succinate (TPGS). Typical micelle formulations including tocopherol-modified therapeutic drug compounds are described in Example 15.

Micelle formulations may also include additional components, such as co-solvents, including the above co-solvents. In one embodiment, the micelle formulation comprises polyethylene glycol and a lower alkyl alcohol (e.g., ethanol). In one embodiment, co-solvents are present in an amount of from about 2 to about 20 weight percent of the total weight of the formulation. The formulations of micelles, emulsions and microemulsions include the aqueous phase. In one embodiment, the aqueous phase comprises deionized water. In another embodiment, the aqueous phase comprises brine. In another embodiment, the aqueous phase is a saline buffered with an organic acid (e.g., succinate, citrate).

The invention also provides the use of compounds of the invention in the manufacture of medicines. For example, for a compound of the invention that includes a therapeutic drug moiety derived from a therapeutic drug compound effective in the treatment of proliferative cell disease, the invention provides the use of such compounds in the manufacture of a medicament for the treatment of proliferative cell disease.

Other aspects also provide methods of administering a compound of the invention to a subject in need thereof and methods of treating a condition that is to be treated by administering a therapeutically effective amount of a compound of the invention. These methods include the administration of the compounds, compositions, emulsion formulations, microemulsion formulations, and micelle formulations described herein.

In one embodiment, the invention provides a method of treating a condition that is to be treated with a parent, unmodified therapeutic drug compound (for example, a proliferative cell disease, such as cancer). In this method, a therapeutically effective amount of a compound of the invention is applied to an object in need thereof.

In one embodiment, the invention provides a method of treating a proliferative cell disease by administering a compound of the invention having a therapeutic drug moiety derived from a therapeutic drug effective in treating a proliferative cell disease. Typical proliferative cell diseases to be treated with the compounds of the invention include hematologic cancers such as leukemia, lymphoma and myeloma; and non-hematologic cancers such as solid tumor carcinomas (e.g., breast, ovary, duodenum, colon, rectum, large lung and bladder cells), sarcomas and gliomas.

Therapeutically effective amounts of these compounds usually vary up to the maximum permissible dose, but these concentrations are not critical and can vary widely. The exact amounts prescribed by the attending physician vary, of course, depending on the compound, route of administration, physical condition of the patient and other factors. The daily dose can be used as a single dose or it can be divided into many doses for administration.

The actually used amount of this compound is a therapeutically effective amount, a term used herein to mean the amount necessary to obtain a significant beneficial effect. Effective doses can be extrapolated from dose-response curves obtained from in vitro or animal model test systems. Animal models are also commonly used to determine the desired dose range and route of administration. Such information can then be used to determine acceptable doses and routes of administration for humans and other mammals. The determination of an effective dose is understood by those skilled in the art. Thus, the amount actually administered depends on the individual being treated, and is preferably the optimal amount such that the desired effect is achieved without noticeable side effects.

The therapeutic efficacy and possible toxicity of a compound of the invention can be determined by standard pharmaceutical procedures in cell cultures or experimental animals (e.g., ED ₅₀ , therapeutically effective dose for 50% of the population; and LD ₅₀ , lethal dose for 50% of the population). The dose ratio between therapeutic and toxic effects is the therapeutic index and can be expressed as the ratio of LD ₅₀ to ED ₅₀ . Modified therapeutic drug compounds that have large therapeutic indices are particularly useful in the practice of the methods of the invention. Data from cell culture and animal studies can be used to determine the dose range for use in humans and other mammals. The dosage of such compounds preferably lies within a range of concentrations that include the ED ₅₀ with little or no toxicity. The dosage usually varies within the indicated range depending on the dosage of the form used, the sensitivity of the patient and the route of administration. Thus, the optimal amounts will vary with the route of administration and are usually in accordance with the amounts of conventional medications administered in the same or similar form.

The compounds of the invention may be administered alone or in combination with one or more additional therapeutic agents. For example, in the treatment of cancer, these compounds can be administered in combination with therapeutic agents, including, but not limited to, androgenic inhibitors such as flutamide and laprolide; antiestrogens such as tomoxifen; antimetabolites and cytotoxic agents, such as daunorubicin, fluorouracil, phloxuridine, alpha interferon, methotrexate, plicamycin, mecaptopurin, thioguanine, adriamycin, carmustine, lomustine, cytarabimucinfucinum, hydroxymorphinustin, hydroxytramubin aminobisamine mitoxantrone, carboplatin, cisplatin, streptozorin, bleomycin, dactinomycin and idamycin; hormones such as medroxyprogesterone, estramustine, ethinyl estradiol, estradiol, leuprolide, megestrol, octreotide, diethylstilbestrol, chlorotrianisene, etoposide, podophyllotoxin and goserelin; nitrogen mustard derivatives such as melphalan, chlorambucil, metlorethamine and thiotepa; steroids such as betamethasone; and other antitumor agents, such as live Mycobacterium bovis, dicarbazine, asparaginase, leucovorin, mitotan, vincristine, vinblastine and taxotere. The respective amounts in each case will vary with the particular agent and are either well known to those skilled in the art or are easily determined using routine experiments.

The administration of the compounds of the invention is carried out in any effective manner, for example, parenterally, topically or orally. Methods of administration include inhalation, buccal, intracerebral, intravenous, intranasal, intrarectal, intraocular, intraarticular, intracervical, intracranial, intracranial, intracranial, intracranial, intracranial, intraocular, , intravertebral, intrathoracic, intratracheal, intra-drum, intrauterine, intravascular and utrizheludochkovoe introduction and other conventional methods. Compounds of the invention having antitumor activity can be administered directly into the tumor, in the vicinity of the tumor, or into a blood vessel that supplies blood to the tumor.

The formulations of the emulsions, microemulsions and micelles of the invention can be sprayed using aerosol propellants, which are known in the field of pulmonary delivery of compounds.

From the compounds of the invention, compositions can be formulated which further comprise suitable pharmaceutically acceptable carriers, including excipients and other compounds that facilitate the administration of the compound to the subject. Further details of the preparation and administration technologies can be found in the latest edition of Remington's Pharmaceutical Sciences (Maack Publishing Co., Easton, PA).

Compositions for oral administration can be prepared using pharmaceutically acceptable carriers well known in the art at a dosage suitable for oral administration. Such carriers allow the preparation of compositions containing the compounds of the invention in the form of tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions suitable for administration by an object. Compositions for oral administration can be prepared, for example, in combination with solid excipients, possibly mixing the resulting mixture and preparing a mixture of granules after adding suitable additional compounds, if necessary, to obtain tablets or dragee cores. Suitable excipients include carbohydrate or protein excipients. These include, but are not limited to, sugars, including lactose, sucrose, mannitol or sorbitol, starch from corn, wheat, rice, potatoes or other plants; cellulose such as methyl cellulose, hydroxypropyl methyl cellulose or sodium carboxymethyl cellulose; and resins, including gum arabic and tragacanth; as well as proteins such as gelatin and collagen. If desired, disintegrating or dissolving agents, such as crosslinked polyvinylpyrrolidone, agar, alginic acid or its salts, such as sodium alginate, can be added.

Dragee cores are provided with suitable coatings, such as concentrated sugar solutions, which may contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol and / or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. Dyes or pigments can be added to tablets or dragee coatings to identify the product or indicate the amount of active ingredient (i.e. dosage).

Compounds for oral administration can be formulated, for example, in the form of tight capsules made of gelatin, or in the form of soft, sealed capsules made of gelatin and a coating such as glycerol or sorbitol. Tight capsules may contain compounds mixed with a filler or a binder, such as lactose or starches, lubricants, such as talc or magnesium stearate, and possibly stabilizers. In soft capsules, covalent compounds can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin or liquid polyethylene glycol, with or without stabilizers.

For topical or nasal administration, penetrating agents are usually used in the formulation corresponding to the particular barrier to be overcome. Examples thereof are 2-pyrrolidone, N-methyl-2-pyrrolidone, dimethylacetamide, dimethylformamide, propylene glycol, methyl or isopropyl alcohol, dimethyl sulfoxide and ozone. Additional agents may be further included to make the formulation cosmetically acceptable. Examples thereof are fats, waxes, oils, paints, flavorings, preservatives, stabilizers and surfactants. Keratolytic agents, such as agents known in the art, may also be included. Examples are salicylic acid and sulfur. For topical administration, the composition may be in the form of a transdermal ointment or dressing for systemically delivering the compound and may be prepared in the usual manner (see, for example, Barry, Dermatological Formulations (Drugs and the Pharmaceutical Sciences-Dekker); Harry's Cosmeticology (Leonard Hill Books).

For rectal administration, the compositions can be used in the form of suppositories or retention enemas. Such compositions can be prepared by mixing the compounds with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid at rectal temperature and therefore melts in the rectum to release the drug. Suitable excipients include, but are not limited to, cocoa butter and polyethylene glycols.

Amounts of each of these various types of additives are readily apparent to those skilled in the art, with optimal amounts being the same as other known formulations formulated for the same type of administration.

Compositions containing the compounds of the invention can be prepared by methods similar to those known in the art (for example, by conventional mixing, dissolving, granulating, dragee-making, powdering, emulsification, encapsulation, inclusion, or lyophilization). The compositions can also be modified to provide appropriate release, delayed release, or target release characteristics by conventional methods (eg, coating). As indicated above, in one embodiment, the compounds are formulated as an emulsion.

Compositions containing the compounds can be prepared in the form of a salt and can be formed with many acids, including, but not limited to, hydrochloric, sulfuric, acetic, lactic, tartaric, malic and succinic. Salts tend to be more soluble in aqueous and other protic solvents than the corresponding free base forms.

Once compositions designed to contain a compound and an acceptable carrier are prepared, they can be placed in an appropriate container and labeled for use. Thus, in another aspect, the invention provides kits.

The tocopherol-modified therapeutic drug compounds of the invention are suitable for administration in both oil-in-water emulsions and micelle formulations. The compounds provide a high load of the drug substance, which makes it possible to administer small volumes.

Emulsions containing tocopherol-modified camptothecin compounds of the invention provide increased stability of the lactone compounds compared to conventional methods for administering camptothecin compounds. A long plasma half-life is achieved for tocopherol-modified camptothecin compounds due to the prolonged effect of the compound on the tumor. Tocopherol-modified compounds achieve high penetration through the lipoid membranes of tumor cells. A greater antitumor effect without increasing toxicity can be achieved by tocopherol-modified camptothecin compounds of the invention as compared to unmodified camptothecin and currently available captecine analogues.

Although the compounds of the invention having formula (2) with m = 1, formula (3) with n = 1 or formula (8) do not include compounds that are not specifically tolerated as described above, it should be appreciated that the compositions, formulation of emulsions, microemulsion formulations and micelle formulations include compounds of the invention having formulas (1) to (8) without similar restrictions. Methods for administering compositions, emulsion formulations, microemulsion formulations and micelle formulations, and methods for preparing the conditions for a person to be treated when administering compositions, emulsion formulations, microemulsion formulations and micelle formulations are also not limited to the compounds of the invention.

The following examples illustrate but do not limit the invention.

EXAMPLES

EXAMPLE 1

Preparation of a typical tocopherol-modified camptothecin compound: camptothecin tocopherol succinate

A 500 ml flask was charged with 10.6 grams of d-α-tocopherol succinic acid, 6.9 grams of camptothecin, 6.13 grams of 2-chloro-1-methylpyridinium iodide (ChMPI), 5.86 grams of 4- (dimethylamino) pyridine ( DMAP) and 200 ml of dry N, N-dimethylacetamide. The mixture was stirred at room temperature for 24 hours and then heated at 50 ° C. for 4 hours. The mixture was cooled to room temperature and then filtered to remove a precipitate and the filtrate was collected. 250 ml of chloroform and 150 ml of deionized water were added to the filtrate to extract the product into chloroform, and the aqueous fraction was removed using a separatory funnel. The chloroform fraction was washed with deionized water (3 × 150 ml) in a separatory funnel, collected and dried over anhydrous MgSO ₄ overnight. MgSO ₄ was removed by filtration, and chloroform was removed using a rotary evaporator under reduced pressure to give a dark yellow solid. The product was purified by silica gel column chromatography. (Yield: 9.5 grams, 55.2%).

¹ H NMR (300 MHz, CDCl ₃ ): δ 8.318 (s, 1H), 8.163-8.135 (d, J = 8.4 Hz, 1H), 7.927-7.901 (d, J = 7.8 Hz, 1H) , 7.842-7.787 (m, 1H), 7.682-7.632 (m, 1H), 7.263-7.242 (d, J = 6.3 Hz, 1H), 5.702-5.410 (Abq, J ₁ = 17.4 Hz, J ₂ = 70 Hz, 2H), 5.190 (s, 2H), 3.014-2.938 (m, 4H), 2.368-0.809 (m, 54H).

Elemental analysis. Calculated for C ₅₃ H ₆₈ N ₂ O ₈ : C, 73.92; H, 7.96; N, 3.25. Found C, 73.61; H, 7.90; N, 3.17.

EXAMPLE 2

Preparation of a typical tocopherol-modified camptothecin compound: 10-hydroxycamptothecin tocopherol succinate

Method 1. Into a 100 ml flask was charged 1.06 grams of d-α-tocopherol succinic acid, 0.476 grams of thionyl chloride and 50 ml of toluene. The mixture was stirred at room temperature overnight. The solvent was removed using a rotary evaporator at 50 ° C and the residue was collected. 0.728 grams of 10-hydroxycamptothecin and 40 ml of dry tetrahydrofuran were added to the residue with stirring. Then, 0.404 grams of triethylamine in 10 ml of tetrahydrofuran was added dropwise to the reaction mixture. The mixture was stirred at room temperature overnight. The mixture was filtered and the white powder was washed with ethyl acetate (3 × 10 ml). The filtrate was collected. The solvent was removed using a rotary evaporator. The residue was collected and purified by column chromatography on silica gel with a mobile phase consisting of acetone and chloroform (1: 4, v / v). (Yield: 0.85 grams, 48.4%).

MS (ES positive ionization): m / z 877 (M) ⁺ .

Analysis. Calculated for C ₅₃ H ₆₈ N ₂ O ₉ : C, 72.58; H, 7.81; N, 3.19. Found: C, 72.52; H, 7.84; N, 3.21.

Alternatively, 10-hydroxycamptothecin tocopherol succinate can be prepared as described below.

Method 2. 2.65 grams of d-α-tocopherol succinate, 0.89 g of thionyl chloride and 20 ml of toluene were charged into a 100 ml flask. The mixture was stirred at room temperature for 24 hours. Toluene and excess thionyl chloride were removed by vacuum distillation at 50 ° C. The residue was dissolved in 15 ml of chloromethane to obtain solution A. 0.9 grams of 10-hydroxycamptothecin, 0.5 ml of triethylamine and 25 ml of freshly dried N, N-dimethylacetamide were added to a 100 ml flask with stirring. Then 15 ml of solution A was slowly added to the mixture through a dropping funnel over 5 minutes. The reaction mixture was stirred at room temperature for 24 hours. The mixture was concentrated by vacuum distillation. 150 ml of ethyl acetate was added to the residue. The mixture was washed with saturated aqueous NaCl (3 × 100 ml). The mixture was dried over anhydrous MgSO ₄ . MgSO ₄ was removed by filtration, and then ethyl acetate was removed by vacuum distillation. The crude product was purified by silica gel column chromatography. (Yield: 1.14 grams, 52.5%).

EXAMPLE 3

Preparation of a typical tocopherol-modified camptothecin compound: 7-ethyl-10-hydroxycamptothecin tocopherol succinate

Method 1: A 500 ml flask was charged with 22.5 grams of d-α-tocopherol succinate, 7.6 g of thionyl chloride and 200 ml of toluene. The mixture was stirred at room temperature for 24 hours. Toluene and excess thionyl chloride were removed by vacuum distillation. The residue was dissolved in 100 ml of chloromethane to obtain solution A. Solution A was used immediately and was not exposed to air. 7.8 grams of 7-ethyl-10-hydroxycamptothecin, 7 ml of triethylamine and 250 ml of freshly dried N, N-dimethylacetamide were loaded into a 500 ml flask with stirring. 100 ml of solution A was slowly added to the mixture through a dropping funnel over 30 minutes. The reaction mixture was stirred at room temperature for 24 hours. The mixture was concentrated by vacuum distillation. 500 ml of ethyl acetate was added to the residue. The mixture was washed with saturated aqueous NaCl (3 × 200 ml). The mixture was dried over anhydrous MgSO ₄ . MgSO ₄ was removed by filtration, and then ethyl acetate was removed by vacuum distillation. The crude product was purified by recrystallization from acetone. (Yield: 15.18 grams, 83.9%).

Melting point 171-173 ° C.

¹ H NMR (300 MHz, CDCl ₃ ): δ 8.236-8.206 (d, J = 9 Hz, 1H), 7.809-7.801 (d, J = 2.4 Hz, 1H), 7.648 (s, 1H), 7.572 -7.533 (dd, J ₁ = 2.7 Hz, J ₂ = 9.3 Hz, 1H), 5.781-5.280 (Abq, J ₁ = 16.2 Hz, J ₂ = 134.0 Hz, 2H), 5.253 (s, 2H), 3.863 (s, 1H), 3.136-3.113 (m, 6H), 2.588 (t, 2H), 2.091 (s, 3H), 2.037 (s, 3H), 1.994 (s, 3H), 1.970-1.852 (m, 2H), 1.821-1.725 (m, 2H), 1.654-0.833 (m, 42H).

MS (ES-positive ionization): m / z 905 (M) ⁺ , 928 (M + Na) ⁺ .

Analysis. Calculated for C ₅₅ H ₇₂ N ₂ O ₉ : C 72.98; H 8.02; N, 3.09.

Found: C, 72.87; H 8.01; N, 2.88.

Alternatively, 7-ethyl-10-hydroxycamptothecin tocopherol succinate can be prepared as described below.

Method 2: 8.48 grams of d-α-tocopherol succinate, 3.81 grams of thionyl chloride and 250 ml of toluene were charged into a 500 ml flask. The mixture was stirred at room temperature overnight. Toluene and excess thionyl chloride were removed using a rotary evaporator at 50 ° C and the residue was collected. 6.27 grams of 7-ethyl-10-hydroxycamptothecin and 250 ml of sodium-dried tetrahydrofuran were added to the residue with stirring. Next, 3.23 grams of triethylamine in 50 ml of tetrahydrofuran was added dropwise to the mixture. The mixture was stirred at room temperature overnight. The mixture was filtered and the white powder was washed with ethyl acetate (3 × 50 ml). The filtrate was collected. The solvent was removed using a rotary evaporator. The crude product was purified by recrystallization in acetone. (Yield: 8.28 grams, 57.2%).

EXAMPLE 4

Obtaining a typical tocopherol-modified compound camptothecin: 10,20-di (tocopherol succinate) 7-ethyl-10-hydroxycamptothecin

Into a 100 ml flask were charged 0.905 grams of 7-ethyl-10-hydroxycamptothecin tocopherol succinate, 0.53 grams of d-α-tocopherolosuccinic acid, 0.255 grams of 2-chloro-1-methylpyridinium iodide, 0.244 grams of 4- (dimethylamino) pyridine and 50 ml of dioxane . The mixture was stirred at room temperature for 24 hours. Thin layer chromatography showed that the reaction had passed. The mixture was filtered to remove a solid phase and the filtrate was collected. The solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography with 30% ethyl acetate in cyclohexane. (Yield: 0.64 grams, 44.82%).

¹ H NMR (300 MHz, CDCl ₃ ): δ 8.168-8.138 (d, J = 9 Hz, 1H), 7.813-7.805 (d, J = 2.4 Hz, 1H), 7.754-7.536 (dd, J ₁ = 2.1 Hz, J ₂ = 11.4 Hz, 1H), 7.197 (s, 1H), 5.703-5.409 (Abq, J ₁ = 17.4 Hz, J ₂ = 71.0, 2H), 5.243- 5.088 (m, 2H), 3.113-2.857 (m, 10H), 2.606-2.564 (t, J = 6 Hz, 2H), 2.383-2.184 (m, 2H), 2.090-1.723 (m, 22H), 1.588- 0.785 (m, 80H).

MS (ES-positive ionization): m / z 1418 (M + H) ⁺ .

Elemental analysis. Calculated for C ₈₈ H ₁₂₄ N ₂ O ₁₃ : C 74.54; H 8.81; N 1.98. Found: C, 74.31; H 8.96; N 1.75.

IR ν _max KBrcm ^-1 : 2925, 2867, 1751, 1665, 1615, 1657, 1510, 1458, 1413, 1376, 1330, 1218, 1128, 1075, 1060, 1035, 992, 943, 923, 829, 812, 758 , 724, 668.

EXAMPLE 5

Preparation of a typical tocopherol-modified camptothecin compound: tocopherol-camptothecin coupled with a hexa (ethylene glycol) binder

Obtaining hexa (ethylene glycol) tocopherol succinate. 2.65 grams of d-α-tocopherol succinic acid and 2.82 grams of hex (ethylene glycol) in 100 ml of toluene were dissolved in a 250 ml flask with stirring. Toluene was removed using a rotary evaporator (drying by azeotropic distillation). 100 ml of chloroform, 1.08 grams of N, N-dicyclohexylcarbodiimide and 100 mg of 4- (dimethylamino) pyridine were added to the mixture. The mixture was stirred overnight. Thin layer chromatography with 40% acetone in hexane showed that the reaction was complete. The mixture was washed three times with deionized water (3 · 100 ml), the chloroform fraction was collected and dried over anhydrous MgSO ₄ for two hours. After filtration, chloroform was removed using a rotary evaporator. The crude product was purified by silica gel column chromatography using successively solvents of 30% ethyl acetate in hexane, 50% ethyl acetate in hexane, and 30% acetone in hexane. (Yield: 0.53 grams, 13.33%).

Obtaining tocopherolsuccinylhexa (ethylene glycol) succinic acid. 1.42 grams of hexa (ethylene glycol) tocopherol succinate obtained above, 0.2 grams of succinic anhydride, 2 drops of tin (II) ethylhexanoate and 25 ml of xylene were charged into a 100 ml flask. The mixture was refluxed for 4 hours. After completion of the reaction, the solvent was removed using a rotary evaporator. The crude product was purified by silica gel column chromatography. (Yield: 0.864 grams, 54%).

Obtaining tocopherol succinate camptothecin with hexa (ethylene glycol) binder. 0.822 grams of tocopherolsuccinylhexa (ethylene glycol) succinic acid obtained above, 0.3 grams of camptothecin, 0.47 grams of 2-chloro-1-methylpyridinium iodide, 0.45 grams of 4- (dimethylamino) pyridine and 40 ml of dry N, N-dimethylacetamide. The reaction mixture was stirred at room temperature overnight. After completion of the reaction, the solvent was removed by vacuum distillation and the residue was collected. 100 ml of ethyl acetate was added to the residue. After stirring for 30 minutes, the mixture was filtered to remove a precipitate, and the filtrate was collected and concentrated. The crude product was purified by silica gel column chromatography. (Yield: 0.342 grams, 30.4%).

¹ H NMR (300 MHz, CDCl ₃ ): δ 8.388 (s, 1H), 8.240-8.213 (d, J = 8.1 Hz, 1H), 7.951-7.923 (d, J = 8.4 Hz, 1H) , 7.861-7.805 (dt, J ₁ = 1.5 Hz, J ₂ = 8.4 Hz, 1H), 7.694-7.640 (dt, J ₁ = 1.2 Hz, J ₂ = 8.1 Hz, 1 H) , 7.269 (s, 1H), 5.708-5.365 (Abq, J ₁ = 17.1 Hz, J ₂ = 85.8 Hz, 2H), 5.283 (s, 2H), 4.273-4.169 (m, 4H), 3.705 -3.673 (t, 2H), 3.631-3.550 (m, 18H), 2.926-2.654 (m, 8H), 2.597-2.552 (t, J = 6.9 Hz, 2H), 2.381-2.113 (m, 2H) , 2.074 (s, 3H), 2.004 (s, 3H), 1.964 (s, 3H), 1.827-1.683 (m, 2H), 1.655 (s, 3H), 1.544-0.964 (m, 24H), 0.875-0.830 (m, 12H).

MS (ES-positive ionization) m / z 1225 (M) ⁺ .

Elemental analysis. Calculated for C ₈₈ H ₁₂₄ N ₂ O ₁₃ : C 67.62; H 7.90; N, 2.29. Found C, 67.08; H 8.04; N, 2.07.

IR ν _max KBr cm ^-1 : 2925, 2867, 1735, 1667, 1618, 1563, 1500, 1457, 1405, 1366, 1349, 1232, 1204, 1141, 1107, 1060, 994, 945, 859, 813, 787, 761, 723, 707.

EXAMPLE 6

Preparation of a typical tocopherol-modified paclitaxel compound: paclitaxel tocopherol succinate

5.83 grams of tocopherol succinic acid, 2.38 grams of thionyl chloride and 50 ml of toluene were charged into a 250 ml flask. The mixture was stirred at room temperature overnight. The solvent was removed using a rotary evaporator at 50 ° C and the residue was collected. 8.54 grams of paclitaxel and 100 ml of dry tetrahydrofuran were added to the residue with stirring. Next, 1.52 grams of triethylamine in 50 ml of tetrahydrofuran was added dropwise to the reaction mixture. The mixture was stirred at room temperature overnight. The mixture was filtered and the white powder was washed with ethyl acetate (3 × 10 ml). The filtrate was collected. The solvent was removed using a rotary evaporator. The precipitate was collected and purified by recrystallization in acetone and hexane. (Yield: 11.56 grams, 84.6%).

Analysis. Calculated for C ₈₀ H ₁₀₃ NO ₁₈ : C, 70.31; H, 7.59; N, 1.02. Found: C, 70.02; H, 7.83; N, 0.93.

EXAMPLE 7

Preparation of a typical tocopherol-modified docetaxel compound: docetaxel tocopherol succinate

A 250 ml flask was charged with 9.86 grams of d-α-tocopherolosuccinic acid, 5.0 grams of docetaxel, 3.83 grams of dry N, N-dicyclohexylcarbodiimide, 500 mg of 4- (dimethylamino) pyridine and 150 ml of chloroform. The mixture was stirred at room temperature overnight. The mixture was filtered to remove a precipitate and the filtrate was collected. The solvent was removed using a rotary evaporator and the residue was collected. The crude product was purified by silica gel column chromatography.

EXAMPLE 8

Obtaining mono-tocopherolphthalate

8.61 grams of dl-α-tocopherol, 2.96 grams of phthalic anhydride, 50 mg of tin (II) 2-ethylhexanoate and 50 ml of dry N, N-dimethylacetamide were charged into a 100 ml flask. The mixture was stirred at about 140 ° C for 24 hours. After cooling the mixture to room temperature, the mixture was poured into 150 ml of ethyl acetate. The mixture was washed three times with saturated aqueous NaCl (3 × 100 ml) and dried over anhydrous MgSO ₄ overnight. The crude product was purified by silica gel column chromatography with 30% ethyl acetate in hexane. (Yield: 3.6 grams, 31.1%).

¹ H NMR (300 MHz, CDCl ₃ ): δ ppm 10.80 (br s, 1H), 8.119-8.063 (m, 1H), 7.883-7.828 (m, 1H), 7.678-7.616 (m, 2H) , 2.627-2.582 (t, 2H), 2.123 (s, 3H), 2.112 (s, 3H), 2.081 (s, 3H), 1.868-1.702 (m, 2H), 1.616-1.020 (m, 24H), 0.874 -0.834 (m, 12H).

Analysis. Calculated for C ₃₇ H ₅₄ O ₅ : C, 76.78; H, 9.40. Found: C, 76.57; H, 9.29.

IR ν _max KBr cm ^-1 : 3073, 2919, 2858, 1737, 1701, 1578, 1455, 1409, 1373, 1276, 1230, 1107, 1071, 913, 738.

EXAMPLE 9

Obtaining mono-tocopherol terephthalate

A 100 ml flask was charged with 4.30 grams of dl-α-tocopherol, 3.32 grams of terephthalic acid, 2.55 grams of 2-chloro-1-methylpyridinium iodide, 0.244 grams of 4- (dimethylamino) pyridine and 50 ml of dry N, N dimethylacetamide. The mixture was stirred at 50 ° C for 4 hours. Thin layer chromatography showed that the reaction was complete. After cooling the mixture to room temperature, the mixture was poured into 150 ml of ethyl acetate. The mixture was washed three times with saturated aqueous NaCl (3 × 100 ml) and dried over anhydrous MgSO ₄ overnight. The crude product was purified by silica gel column chromatography with 30% ethyl ether in hexane. (Yield: 1.60 grams, 27.6%).

¹ H NMR (300 MHz, CDCl ₃ ): δ ppm 11.80 (br s, 1H), 8.374-8.259 (q, J ₁ = 8.4 Hz, J ₂ = 26.1 Hz, 4H), 2.650 -2.607 (t, 2H), 2.130 (s, 3H), 2.066 (s, 3H), 2.024 (s, 3H), 1.895-1.783 (m, 2H), 1.532-1.083 (m, 24H), 0.878-0.839 (m, 12H).

Analysis. Calculated for C ₃₇ H ₅₄ O ₅ : C 76.78; H, 9.40. Found: C 76.64; H, 9.39.

IR ν _max KBr cm ^-1 : 3062, 2924, 2858, 1737, 1696, 1573, 1460, 1424, 1373, 1276, 1240, 1097, 928, 774, 723.

EXAMPLE 10

Preparation of a typical tocopherol-modified camptothecin compound: camptothecin tocopherol terephthalate

1.16 grams of the mono-tocopherol prepared above, 0.70 grams of camptothecin, 0.511 grams of 2-chloro-1-methylpyridinium iodide and 0.489 grams of 4- (dimethylamino) pyridine were charged into a 100 ml flask. The mixture was stirred at 50 ° C. overnight. Thin layer chromatography showed that the reaction was complete. After cooling the mixture to room temperature, the reaction mixture was poured into 150 ml of ethyl acetate. The mixture was filtered and the filtrate was collected. The filtrate was washed with saturated aqueous NaCl (3 × 100 ml) and dried over anhydrous MgSO ₄ overnight. The crude product was purified by silica gel column chromatography. (Yield: 0.560 grams, 30.8%).

¹ H NMR (300 MHz, CDCl ₃ ): δ ppm 8.387 (s, 1H), 8.370-8.242 (q, J ₁ = 8.4 Hz, J ₂ = 30.3 Hz, 4H), 8.167-8.139 (d , J = 8.4 Hz, 1H), 7.937-7.910 (d, J = 8.1 Hz, 1H), 7.823-7.774 (t, 1H), 7.672-7.625 (t, 1H), 7.260 (s, 1H) ), 5.823-5.462 (Abq, J ₁ = 17.4 Hz, J ₂ = 90.9 Hz, 2H), 5.302 (s, 2H), 2.461 (t, 2H), 2.559-2.312 (m, 2H), 2.123 (s, 3H), 2.056 (s, 3H), 2.015 (s, 3H), 1.844-1.801 (m, 2H), 1.629-1.085 (m, 27H), 0.938-0.789 (m, 12H).

Analysis. Calculated for C ₅₇ H ₆₈ N ₂ O ₈ : C 75.30; H 7.54; N, 3.08. Found: C, 74.91; H 7.56; N, 3.02.

IR ν _max ^KBr cm ^-1 : 3057, 2924, 2858, 1757, 1737, 1675, 1614, 1558, 1450, 1399, 1266, 1235, 1163, 1102, 1020, 723.

EXAMPLE 11

Preparation of a typical tocopherol-modified camptothecin compound: tocopherol-cyclohexane-1,2-dicarboxylate 7-ethyl-10-hydroxycamptothecin

Preparation of tocopherol cyclohexane-1,2-dicarboxylic acid. A mixture of 1.54 grams of 1,2-cyclohexanedicarboxylic acid anhydride, 8.6 grams of d-α-tocopherol, 1.34 grams of aluminum trichloride and 100 ml of cyclohexane was refluxed in a 250 ml flask for about 30 minutes. After the mixture was cooled to room temperature, it was filtered. The filtrate was washed with dilute aqueous hydrochloric acid and then dried over anhydrous MgSO ₄ . The mixture was concentrated and the crude product was purified by silica gel column chromatography. (Yield: 3.325 grams, 56.9%).

Preparation of tocopherol cyclohexane-1,2-dicarboxylate of 7-ethyl-10-camptothecin. A mixture of 1.08 grams of tocopherol cyclohexane-1,2-dicarboxylic acid prepared above, 0.44 grams of thionyl chloride and 20 ml of toluene was stirred under nitrogen overnight. Toluene and excess thionyl chloride were removed by vacuum distillation and the residue was dissolved in 10 ml of dichloromethane to obtain a solution A. 0.350 grams of SN38 were dissolved in a 100 ml flask in 25 ml of dry N, N-dimethylacetamide to obtain a solution of B. Solution A and 0.186 g of triethylamine were added to the solution C. The mixture was stirred overnight at room temperature. The crude product was purified by silica gel column chromatography. (Yield 0.59 grams, 68.9%).

EXAMPLE 12

Preparation of a typical tocopherol-modified compound of doxorubicin: doxorubicin tocopherol succinate

Equivalent moles (1 mmol) of tocopherol succinic acid, doxorubicin, N, N-dicyclohexylcarbodiimide and 50 ml of dry N, N-dimethylacetamide were charged into a 100 ml flask. The mixture was stirred at room temperature until completion of the reaction. The mixture was filtered to remove a white precipitate and the filtrate was collected. The solvent was removed using a rotary evaporator and the residue was collected. The product was purified either by recrystallization or column chromatography on silica gel.

EXAMPLE 13

Preparation of a typical tocopherol-modified hydroxyzine compound: hydroxyzine tocopherol succinate

Equivalent moles (1 mmol) of tocopherol succinic acid and thionyl chloride and 50 ml of toluene were charged into a 100 ml flask. The mixture was stirred at room temperature overnight. The solvent was removed using a rotary evaporator at 50 ° C and the residue was collected. 1 mmol of hydroxyzine and 40 ml of chloroform were added to the residue with stirring. Then, 1 mmol of triethylamine in 10 ml of chloroform was added dropwise to the reaction mixture at 0-5 ° C. The mixture was then stirred at room temperature overnight. The mixture was washed with saturated aqueous NaHCO ₃ (3 x 50 ml). The organic phase was collected and dried over anhydrous MgSO ₄ . The solvent was removed using a rotary evaporator after removal of MgSO ₄ . The residue was collected and the crude product was purified either by recrystallization or column chromatography on silica gel.

EXAMPLE 14

The solubility of a typical tocopherol-modified therapeutic drug compound

In this example, the solubility of typical tocopherol-modified therapeutic drug compounds of the invention, camptothecin tocopherol succinate and 7-ethyl-10-hydroxycamptothecin tocopherol succinate were compared with camptothecin solubility in various solvents.

The solubilities of camptothecin, camptothecin tocopherol succinate and 7-ethyl-10-hydroxycamptothecin tocopherol succinate were determined in several solvents. Compounds were dissolved in each solvent with constant stirring and temperature until saturation. The resulting solutions were centrifuged and the supernatant was analyzed by high performance liquid chromatography (HPLC).

The comparative solubility (mg / g) of camptothecin, camptothecin tocopherol succinate and 7-ethyl-10-hydroxycamptothecin tocopherol succinate in various solvents is presented in table 1.

Table 1
Comparison of the solubility of camptothecin and camptothecin tocopherol succinate Solvent Camptothecin
(mg / g) VESA-SN38 ^one
mg / g) VESA-CPT ²
(mg / g) Temperature
(° C) PEG-400
Nf - 0.017 17.5 Room
temperature TPGS - 27.6 > 133.3 65 Vitamin E
USP / NF 1.96 398.2 > 283.3 65 Soybean oil
USP 0.00 3.3 45,2 Room
temperature Captex 300
EP - 4.4 96.7 Room
temperature Twin 80
Nf - 2.7 48.3 65 → Room
temperature Ethanol
denatured alcohol - 4.2 57.1 Room
temperature Methanol - 3.8 14,4 Room
temperature Acetonitrile 0.09 3,1 49.6 Room
temperature Chloroform 0.71 > 97.0 > 372.5 Room
temperature DMSA fifty 30.5 > 255.5 Room
temperature Methylene
dichloride 0.9 > 99.7 > 336.5 Room
temperature Propylene glycol, USP - 3.2 0.4633 Room
temperature Glycerol
USP / EP / BP /
JP - 2,8 2,541 Room
temperature 1 VESA-SN38: 7-ethyl-10-hydroxycamptothecin tocopherol succinate 2 VESA-CPT: tocopherol succinate camptothecin

The results in Table 1 illustrate that camptothecin tocopherol succinate and 7-ethyl-10-hydroxycamptothecin tocopherol succinate both have significant oil solubility and have particularly high solubility in vitamin E (α-tocopherol).

EXAMPLE 15

Emulsions containing typical tocopherol-modified therapeutic drug compounds

This example describes typical emulsions containing tocopherol-modified therapeutic drug compounds.

A. Emulsion of 7-ethyl-10-hydroxycamptothecin tocopherol succinate.

Tocopherol succinate 7-ethyl-10-hydroxycamptothecin, prepared as described in Example 3, was dissolved in vitamin E and then emulsified using a microfluidizer (M110Y Microfluidics) in the presence of d-α-tocopherol polyethylene glycol 1000 succinate (TPGS), poloxamer 407 and c an emulsion having the following composition (wt.%):

7-ethyl-10-hydroxycamptothecin tocopherol succinate 0.69% Vitamin E 7.31% TPGS 5% Poloxamer 407 one% Saline solution 86%

The emulsion was filtered through a 0.2 μm filter and poured into sterile glass vials. The average particle size was approximately 50 nM, as determined by a submicron size sizer (Nicomp Model 370), with 99% of the particles smaller than 80 nm. No signs of precipitate formation or decrease in concentration when measured by HPLC were observed for at least 3 months when stored at 4 ° C.

B. Emulsion of tocopherol succinate 7-ethyl-10-hydroxycamptothecin.

Tocopherol succinate 7-ethyl-10-hydroxycamptothecin obtained as described in Example 3 was dissolved in vitamin E and then emulsified using a microfluidizer (M110Y Microfluidics) in the presence of TPGS and saline to obtain an emulsion having the following composition (wt.%):

7-ethyl-10-hydroxycamptothecin tocopherol succinate 0.69% Vitamin E 7.31% TPGS 5% Saline solution 87%

This formulation formed a more yellow and coarse emulsion than the emulsion obtained as described above, which included Poloxamer 407. The emulsion was filtered through a 0.2 μm filter and poured into sterile glass vials. The average particle size was approximately 75 nm, as determined by a submicron size sizer (Nicomp Model 370), with 99% of the particles smaller than 170 nm. No signs of precipitate formation or decrease in concentration when measured by HPLC were observed for at least 3 months when stored at 4 ° C.

C. Emulsion of tocopherol succinate camptothecin.

Camptothecin tocopherol succinate obtained as described in Example 1 was dissolved in vitamin E and then emulsified using a microfluidizer (M110Y Microfluidics) in the presence of TPGS, poloxamer 407 and saline to obtain an emulsion having the following composition (wt.%):

Camptothecin tocopherol succinate 0.74% Vitamin E 7.26% TPGS 5% Poloxamer 407 one% Saline solution 86%

The emulsion was filtered through a 0.2 μm filter and poured into sterile glass vials. The average particle size was approximately 40 nm, as determined by a submicron size determinant (Nicomp Model 370), with 99% of the particles smaller than 75 nm. No signs of precipitate formation or a decrease in concentration when measuring HPLC were observed for at least 3 months when stored at 4 ° C.

D. Emulsion of tocopherol succinate camptothecin.

Camptothecin tocopherol succinate obtained as described in Example 1 was dissolved in vitamin E and then emulsified using a microfluidizer (M110Y Microfluidics) in the presence of TPGS, poloxamer 407 and saline to obtain an emulsion having the following composition (wt.%):

Tocopherol succinate camptothecin 1.48% Vitamin E 6.52% TPGS 5% Poloxamer 407 one% Saline solution 86%

The emulsion was filtered through a 0.2 μm filter and poured into sterile glass vials. The average particle size was approximately 30 nm, as determined by a submicron size sizer (Nicomp Model 370), with 99% of the particles smaller than 100 nm. No signs of precipitate formation or a decrease in concentration when measuring HPLC were observed for at least 3 months when stored at 4 ° C.

E. Emulsion of tocopherol succinate 7-ethyl-10-hydroxycamptothecin.

The 7-ethyl-10-hydroxycamptothecin tocopherol succinate obtained as described in Example 3 was dissolved in vitamin E and then emulsified using a microfluidizer (M110Y Microfluidics) in the presence of TPGS and citric acid buffered saline to obtain an emulsion having the following composition (wt.% ):

7-ethyl-10-hydroxycamptothecin tocopherol succinate 0.69% Vitamin E 7.31% TPGS 5% Buffer solution of citric acid, pH 3 87%

The emulsion was filtered through a 0.2 μm filter and poured into sterile glass vials. The average particle size was approximately 60 nm, as determined by a submicron size sizer (Nicomp Model 370), with 99% of the particles smaller than 150 nM. No signs of precipitate formation or a decrease in concentration when measuring HPLC were observed for at least 3 months when stored at 4 ° C and 25 ° C.

F. Emulsion of tocopherol succinate 7-ethyl-10-hydroxycamptothecin.

The 7-ethyl-10-hydroxycamptothecin tocopherol succinate obtained as described in Example 3 was dissolved in vitamin E and then emulsified using a microfluidizer (M110Y Microfluidics) in the presence of TPGS and succinate buffered saline to obtain an emulsion having the following composition (wt.%) :

Recipe 1 7-ethyl-10-hydroxycamptothecin tocopherol succinate 0.69% Vitamin E 7.31% TPGS 5% Succinate Buffer Saline, pH 4.0 87%

The emulsion was filtered through a 0.2 μm filter and poured into sterile glass vials. The average particle size was approximately 70 nm, as determined by a submicron size determinant (Nicomp Model 370), with 99% of the particles smaller than 170 nm. No signs of precipitate formation or a decrease in concentration when measuring HPLC were observed for at least 3 months when stored at 4 ° C and 25 ° C.

Recipe 2 7-ethyl-10-hydroxycamptothecin tocopherol succinate one% Vitamin E 7% TPGS 5% Succinate Buffer Saline, pH 4.0 87%

The emulsion was filtered through a 0.2 μm filter and poured into sterile glass vials. The average particle size was approximately 70 nm, as determined by a submicron size determinant (Nicomp Model 370), with 99% of the particles smaller than 170 nm. No signs of precipitate formation or a decrease in concentration when measuring HPLC were observed for at least 1 month when stored at 4 ° C, 25 ° C and 40 ° C.

Recipe 3 7-ethyl-10-hydroxycamptothecin tocopherol succinate one% Vitamin E 6% TPGS four% Succinate Buffer Saline, pH 4.0 89%

The emulsion was filtered through a 0.2 μm filter and poured into sterile glass vials. The average particle size was approximately 95 nm, as determined by a submicron particle size determinant (Nicomp Model 370), with 99% of the particles smaller than 220 nm. No signs of precipitate formation or a decrease in concentration when measuring HPLC were observed for at least 1 month when stored at 4 ° C, 25 ° C and 40 ° C.

G. Formulation of 7-ethyl-10-hydroxycamptothecin tocopherol succinate micelles (VESA-SN38)

7-ethyl-10-hydroxycamptothecin tocopherol succinate was dissolved in a mixture containing TPGS, PEG (300) and ethanol at about 50 ° C. to about 60 ° C. with stirring for about an hour until a clear solution formed. To this solution was added deionized water (DI-water), Poloxamer 407 and DI-water, Poloxamer 188 and DI-water or 0.9% aqueous NaCl solution to form 1-5 formulations, respectively, below. The formulations were mixed for several minutes to form clear micelle solutions having the following compositions (wt.%):

Recipe 1 VESA-SN38 0.2% TPGS 5% Ethanol 5% PEG (300) 5% Di water 84.8%

The solution formulation was filtered through a 0.2 μm filter and poured into sterile glass vials. No evidence of precipitate formation or decrease in concentration was observed by measuring HPLC for at least 11 weeks when stored at 4 ° C.

Recipe 2 VESA-SN38 0.2% TPGS 5% Poloxamer 407 1.7% Ethanol 5% PEG (300) 5% Di water 83.1%

The formulation solution was filtered through a 0.2 μm filter and poured into sterile glass vials. No evidence of precipitate formation or decrease in concentration was observed by measuring HPLC for at least 11 weeks when stored at 4 ° C.

Recipe 3 VESA-SN38 0.2% TPGS 5% PEG (300) 5% Ethanol 5% Poloxamer 188 1.7% Di water 83.1%

The formulation solution was filtered through a 0.2 μm filter and poured into sterile glass vials. No signs of precipitate formation or a decrease in concentration when measuring HPLC were observed for at least 11 weeks when stored at 4 ° C and 25 ° C.

Recipe 4 VESA-SN38 0.2% TPGS 2% PEG (300) 2% Ethanol four% Saline solution 91.8%

The formulation solution was filtered through a 0.2 μm filter and poured into sterile glass vials. No signs of precipitate formation or decrease in concentration when measuring HPLC were observed for at least 1 week when stored at 4 ° C, 25 ° C and 40 ° C.

Recipe 5 VESA-SN38 0.5% TPGS 5% PEG (300) 5% Ethanol 10% Saline solution 79.5%

The formulation solution was filtered through a 0.2 μm filter and poured into sterile glass vials. No evidence of precipitate formation or decrease in concentration was observed by measuring HPLC for at least 3 weeks when stored at 4 ° C.

H. Formulation of 7-ethyl-10-hydroxycamptothecin tocopherol succinate micelles (VESA-SN38)

7-ethyl-10-hydroxycamptothecin tocopherol succinate was dissolved in a mixture containing TPGS, PEG (300) and ethanol at about 50 ° C. to about 60 ° C. with stirring for about 1 hour until a clear solution formed. Succinate buffered saline was added to this solution to formulate Formulations 1 and 2 below. The formulations were mixed for several minutes to form clear micelle solutions having the following compositions (wt.%):

Recipe 1 VESA-SN38 0.2% TPGS 2% Ethanol four% PEG (300) 2% Succinate Buffer Saline, pH 4.0 91.8%

The formulation solution was filtered through a 0.2 μm filter and poured into sterile glass vials.

Recipe 2 VESA-SN38 0.5% TPGS 5% Ethanol 10% PEG (300) 5% Buffered Saline
succinate pH 4.0 79.5%

The formulation solution was filtered through a 0.2 μm filter and poured into sterile glass vials.

EXAMPLE 16

In vitro stability of the lactone of a typical tocopherol-modified therapeutic drug compound in the presence of human albumin

In this in vitro example, the stability in the presence of human albumin of the lactone form of a typical tocopherol-modified therapeutic drug compound of the invention, tocopherol succinate camptothecin and tocopherol succinate 7-ethyl-10-hydroxycamptothecin was compared in vitro with the stability of the lactone form camptothecin.

Since lactone (cycle E) is a critical fragment for camptothecin activity and it is reported to be unstable under physiological conditions (pH 7.4), the stability of camptothecin tocopherol succinate lactone and 7-ethyl-10-hydroxycamptothecin tocopherol succinate was determined. It was assumed that the solubilization of camptothecin tocopherol succinate and 7-ethyl-10-hydroxycamptothecin tocopherol succinate in the oil phase protects the lactone from hydrolysis and, thus, provides improved stability of the lactone under physiological conditions. To assess the stability of lactone, a saline buffer solution (10 mM, pH 7.4) containing 4% human albumin serum was incubated at 37 ° C in the presence of camptothecin (dissolved in DMSO), camptothecin tocopherol succinate emulsion (prepared as described in Example 15C) , designated herein as “SN2300 emulsion”) or 7-ethyl-10-hydroxycamptothecin tocopherol succinate emulsions (prepared as described in Example 15A, designated herein as “SN2310 Emulsion”). High performance liquid chromatography with a fluorescent detector was used to analyze the decrease in concentration of the lactone form over time.

Figure 12 illustrates the percentage change in concentration of the lactone form over time for camptothecin, camptothecin tocopherol succinate (SN2300) and 7-ethyl-10-hydroxycamptothecin tocopherol succinate (SN2310) in the presence of human albumin. The stability of lactone tocopherol succinate camptothecin and tocopherol succinate 7-ethyl-10-hydroxycamptothecin is greater than the stability of camptothecin. This sharp increase in lactone resistance can lead to increased activity compared to the unmodified starting camptothecin compound.

EXAMPLE 17

In vitro cytotoxicity of typical tocopherol-modified therapeutic drug compounds

In this example, the in vitro cytotoxicity of the typical tocopherol-modified therapeutic drug compounds of the invention, captothecin tocopherol succinate and 7-ethyl-10-hydroxycamptothecin tocopherol succinate were compared with the in vitro cytotoxicity of camptothecin, 10-hydroxycaptothecin, SN38, irinotecan and

In vitro cytotoxicity, measured by RI ₅₀ values (50% increase in inhibition) for camptothecin tocopherol succinate and 7-ethyl-10-hydroxycamptothecin tocopherol succinate, was investigated and compared with RI ₅₀ values at the National Cancer Institute (NCI) for camptothecin, 10-hydroxycamptothecin, SN -38, irinotecan and topotecan in the following colonies of cancer cells: NCI-H460 (ATCC # HTB-177) (large lung cells), HCT-15 (ATCC # CCL-225) (rectal), HT-116 (ATCC # CCL- 247) (rectal), HT-29 (ATCC # HTB-38) (rectal), MCF-7 (ATCC # HTB-22) (thoracic) and OVCAR-3 (ATCC # HTB-161) (ovarian).

The study was carried out using the formulations of emulsions of tocopherol succinate 7-ethyl-10-hydroxycamptothecin (described in Example 15A) and camptothecin tocopherol succinate (described in Example 15C), diluted in an appropriate cell medium. Cells were in contact with various concentrations of test compounds for 48 hours. After 48 hours, staining was performed using ALamar Blue to determine the number of viable cells and calculate the level of inhibition of cell growth when compared with the control group. The percentage inhibition of concentration was calculated using the Hill equation to determine the concentration that causes a 50% increase in inhibition (RI ₅₀ ).

The sensitivity of the tested cell lines to camptothecin tocopherol succinate (SN2300) and 7-ethyl-10-hydroxycamptothecin tocopherol succinate (SN2310), camptothecin, irinotecan and topotecan are illustrated in table 2 and figure 13.

table 2
Comparative concentration of the drug, which causes a 50% increase in inhibition (RI _fifty ) The colony
cells CPT
(NCl) 10-HO-
CPT
(NCl) SN38
(NCl) Irinotecan
(NCl) Topotecan
(NCl) VESA-
CPT VESA-
SN38 NCl-H460
(NSCLC) 16 nM 11 nM 1.4 nM 5.01 μM 19.9 nM 43 nM 4 nM HCT-15
(thick
intestines) 160 nM 356 nm 7.9 nM 31.6 μm 501 nM 20 μm 99 nM OVCAR-3
(ovaries) 160 nM 62 nM 2.9 nM 31.6 μm 251 nM Weak
assets-
nost 83 nM HCT-116
(thick
intestines) 40 nM 27 nM 21 nM 7.9 μM 39.8 nM 449 nM 119 nM Ht29
(thick
intestines) 126 nM 112 nM 1 nM 12.58 μM 125 nM 434 nm 91nM Mcf-7
(chest) 13 nM 10 nM 3.98 μM 15.8 nM 325 nM CPT: camptothecin; 10-HO-CPT: 10-hydroxycamptothecin; SN38: 7-ethyl-10-hydroxycamptothecin; VESA-CPT tocopherol succinate camptothecin; VESA-SN38: 7-ethyl-10-hydroxycamptothecin tocopherol succinate.

The results in Table 2 illustrate that the formulations of the tocopherol-modified therapeutic drug compounds of the invention provide effective antitumor activity.

FIGURE 13 is a graph of RI ₅₀ values (a concentration that causes a 50% increase in inhibition) determined for camptothecin tocopherol succinate and 7-ethyl-10-hydroxycamptothecin tocopherol succinate in four tested cell colonies. The values reported by the NCI for the same cell colonies for camptothecin, irinotecan and topotecan are also included for comparison. A high RI ₅₀ value corresponds to a low concentration of the drug, causing 50% inhibition. From the diagram it is clear that the compounds of the invention tocopherol succinate camptothecin and tocopherol succinate 7-ethyl-10-hydroxycamptothecin show a high level of cytotoxic activity, similar to camptothecin.

EXAMPLE 18

Pharmacokinetics of typical tocopherol-modified therapeutic drug compounds

In this example, the pharmacokinetics of typical tocopherol-modified therapeutic drug compounds of the invention tocopherol succinate camptothecin and tocopherol succinate 7-ethyl-10-hydroxycamptothecin are compared with camptothecin, irinotecan and topotecan.

The pharmacokinetic profiles of camptothecin tocopherol succinate and 7-ethyl-10-hydroxycamptothecin tocopherol succinate were studied in Sprague-Dowley rats after pills, intravenous administration of drug emulsion formulations (SN2300 emulsion and SN2310 emulsion) through a lateral tail vein with a dose of about 14 mg, approximately body. Blood samples were taken up to 120 hours after dosing through the jugular vein. The concentration of each camptothecin derivative in plasma was determined by high performance liquid chromatography (HPLC) with a fluorescence detector. General analysis was performed using WinNonlin (v 4.1).

The pharmacokinetic profiles of tocopherol succinate camptothecin (SN2300) and tocopherol succinate 7-ethyl-10-hydroxycamptothecin (SN2310) are shown in FIGURES 14A and 14B, respectively. FIGURES 14A and 14B show concentration-time values after intravenous injection of 13.8 mg of the drug compound / kg body weight for tocopherol succinate camptothecin (SN2300 emulsion) and tocopherol succinate 7-ethyl-10-hydroxycamptothecin (SN2310 emulsion), respectively. With reference to FIGURES 14A and 14B, a prolonged half-cycle of plasma conversion after intravenous administration is shown, especially for camptothecin tocopherol succinate.

The calculated plasma elimination half-life, average retention time and excretion of camptothecin tocopherol succinate (SN2300), 7-ethyl-10-hydroxycamptothecin tocopherol succinate (SN2310), camptothecin, irinotecan and topotecan are presented in Table 3.

The term "plasma elimination half-life" refers to the time required to reduce the concentration of the drug in plasma to 50% after reaching equilibrium. The term “elimination rate constant” refers to the proportion of a drug eliminated per unit time. In the first elimination order, the elimination rate is directly proportional to the concentration of the drug in serum. There is a linear relationship between the elimination rate and the concentration of the drug in serum. Although the amount of drug eliminated in the first order process varies with concentration, the proportion of eliminated drug remains constant.

The term “purification” refers to measuring the body’s ability to excrete a drug and represents the hypothetical volume of drug distribution that is output per unit of time (ie ml / min) by any means of drug removal. It is important to make it clear that the purification does not show how much drug has been removed, but rather the volume of biological fluid, such as blood or plasma, that would have to be free of the drug by excretion. The term "distribution volume" refers to the calculated volume of body fluid that would be required to dissolve the total amount of the drug with the same concentration that is found in the blood. This is a proportionality constant that relates the amount of the drug in the body to the measured concentration in the biological fluid (blood, plasma, serum).

Table 3
Comparative pharmacokinetic parameters after intravenous administration to rats of tocopherol succinate camptothecin (SN2300) and tocopherol succinate 7-ethyl-10-hydroxycamptothecin (SN2310) in comparison with camptothecin, irinotecan and topotecan Compound t _1/2 (hour)
(elimination) MRT
(clock) Cleaning
(l / hour / kg) SN2300 29.08 9.40 0.0081 SN2310 3.49 5.15 0.0067 Camptothecin ^a 1.7 - 8.81 Topotecan 1.06 - 3.22 Irinotecan ^b 1,54 1.43 2.22 ^a El-Gizawy SA, Hedaya MA. Cancer Chemother. Pharmacol., 43: 364-370 (1999). ^b Atsumi R, Okazaji O Hakusui H. Biol. Pharm Bull., 18 (8): 1114-1119 (1995).

Table 3 illustrates that the calculated elimination half-life of camptothecin tocopherol succinate (SN2300) and 7-ethyl-10-hydroxycamptothecin tocopherol succinate (SN2310) elimination is approximately 30 times and 3 times, respectively, more than commercially available analogues. A higher average retention time (IED) and a lower cleaning rate suggests a longer treatment time for the tumor with these new derivatives, which may indicate the possibility of an increase in the chemotherapeutic effect.

Through the lipophilic modification of therapeutic drug compounds, the plasma elimination half-life of the parent therapeutic drug compound can be increased. Compounds according to the invention, due to the advantages of a lipophilic fragment (for example, a tocopherol fragment), have an increased plasma elimination half-life compared to the initial therapeutic drugs. As shown above, for tocopherol succinate camptothecin and tocopherol succinate 7-ethyl-10-hydroxycamptothecin, the plasma elimination half-life is significantly increased in comparison with the starting compounds.

EXAMPLE 19

In vivo antitumor activity of typical tocopherol-modified therapeutic drug compounds in human tumor xenografts

In this in vivo example, the antitumor activity of typical tocopherol-modified therapeutic drug compounds of the invention, tocopherol succinate camptothecin and tocopherol succinate 7-ethyl-10-hydroxycamptothecin are compared with the antitumor effect of irinotecan.

The NCI-H460 xenograft is a human tumor. Atymic mice were implanted subcutaneously with a cell suspension (10 ⁷ cells / mouse). When the tumors reached the appropriate size, the animals were randomized into groups of eight and the following compounds were administered intravenously with a dose of 15 mg of the drug compound / kg body weight according to the q1dx5 regimen for the next two weeks:

Salt control group

Irinotecan

SN2300 emulsion

SN2310 emulsion

HT-29 human tumor xenograft. Atymic mice were subcutaneously implanted with 30-40 mg of tumor fragments using 12-size trocar needles. A sufficient number of mice implanted with tumor fragments in a narrow weight range (100-200 mg) were selected for testing on an installation day (UD). Animals selected with tumors in the appropriate size range were randomized into six groups of 10 animals and were given the following test compounds subcutaneously:

Salt control group (q1dx5 for 2 weeks) Irinotecan (15 mg / kg, q1dx5 for two weeks) SN2300 emulsion (15 mg / kg, q1dx5 for two weeks) SN2310 emulsion (15 mg / kg, q1dx5 for two weeks) SN2300 emulsion (15 mg / kg, q3dx10) SN2310 emulsion (15 mg / kg, q3dx10)

In both xenograft studies, animal body weight and tumors were measured twice a week after dosing. Tumor measurements were performed using a compass (millimeters); tumor volume was calculated based on the formula: (length × width ² ) / 2 = volume (mm ³ ).

The antitumor effect of SN2300 and SN2310 emulsions administered to atimic mice implanted with NCI-H460 human tumor cells and HT-29 human tumor cells is graphically shown in FIGURES 15A and 15B, respectively. Although the SN2300 emulsion showed a weak antitumor effect in this model or its absence, the SN2310 emulsion showed a significant antitumor effect in comparison with both the saline control solution and irinotecan.

The calculated response parameters for the HT-29 xenograft study are presented in Table 4. Fifty-five days after the dose, 30% of the mice in the control group were euthanized due to tumor size (> 4000 mm ³ ) and the average tumor size was 3136 mm ³ . At the same time, 80% of mice in the SN2310 (q3dx10) group had immeasurable tumors. In addition, the SN2310 group (q1dx5) had an average tumor size of 126 mm ³ with 40% having immeasurable tumors. At the same time, in the irinotecan group, the average tumor size was 1637 mm ³ . The results show that the administration of SN2310 induces significant antitumor activity.

Table 4
The calculated tumor response parameters from the study of HT-29 xenograft Group Scheme Tumor growth inhibition (T-C)
(days) Inhibition
growth
tumors
(% T / C) Number
animals with immeasurable tumor
on day 55 Saline solution q1dx5
(2 weeks) ---- ---- 0/10 Irinotecan q1dx5
(2 weeks) fourteen 52 0/10 SN2300 q1dx5
(2 weeks) 8.5 79 0/10 SN2300 q3dx10 0 102 0/10 SN2310 q1dx5
(2 weeks) > 28 ^* four 4/10 SN2310 q3dx10 > 28 ^* 2 8/10 ^* 90% of tumors in this group have not yet reached the maximum of a predetermined size. T-C = average time for the treatment group (T) and the control group (C) to achieve a predetermined size. % T / C = (average weight of the treated tumor) / (average weight of the control tumor) x100 (on day 55).

While a preferred embodiment of the invention has been illustrated and described, it should be borne in mind that various changes can be made without going beyond the scope of the idea and scope of the invention.

Embodiments of the invention in which exclusive ownership or advantage is claimed are defined as follows.

Claims (18)

1. The compound having the formula T-L-D, in which T represents tocopherol, L is succinate, and D is camptothecin or its derivative, where all three fragments are covalently linked. 2. The compound according to claim 1, in which D is 10-hydroxycamptothecin or its derivative. 3. The compound according to claim 1, in which D is 7-ethyl-10-hydroxycamptothecin or its derivative. 4. Tocopherol succinate docetaxel. 5. Tocopherol succinate camptothecin. 6. Tocopherol succinate 10-hydroxycamptothecin. 7. Tocopherol succinate 7-ethyl-10-hydroxycamptothecin. 8. An emulsion for the treatment of cell proliferative disease, containing (a) an oil phase containing (i) a compound having the formula T-L-D, in which T represents tocopherol, L is succinate, and D is camptothecin or its derivative, where all three fragments are covalently linked, (ii) lipophilic medium; and (b) the aqueous phase. 9. The emulsion according to claim 8, in which the lipophilic medium contains tocopherol. 10. The micelle formulation for the treatment of cell proliferative disease, containing (a) a compound having the formula T-L-D, in which T represents tocopherol, L is succinate, and D is camptothecin or its derivative, where all three fragments are covalently linked, and (b) the aqueous phase. 11. A method of treating cell proliferative disease, comprising administering to the subject, if necessary, therapeutically effective amounts of a compound of the formula T-L-D, in which T represents tocopherol, L is succinate, and D is camptothecin or its derivative, where all three fragments are covalently linked. 12. The method according to claim 11, in which the introduction of the compound includes the introduction of an emulsion containing the compound. 13. The method of claim 11, wherein administering the compound comprises administering a micelle formulation containing the compound. 14. The compound having the formula T-L-D, in which T represents tocopherol, L is succinate, and D is docetaxel, where tocopherol is covalently linked to docetaxel via a succinate moiety. 15. An emulsion for the treatment of cell proliferative disease, containing (a) an oil phase containing (i) a compound having the formula T-L-D, in which T represents tocopherol, L is succinate, and D is docetaxel, where tocopherol is covalently linked to docetaxel via a succinate moiety, (ii) lipophilic medium; and (b) the aqueous phase. 16. The emulsion according to clause 15, in which the lipophilic medium contains tocopherol. 17. A method of treating cell proliferative disease, comprising administering to the subject, if necessary, therapeutically effective amounts of a compound of the formula T-L-D, in which T represents tocopherol, L is succinate, and D is docetaxel, where tocopherol is covalently linked to docetaxel via a succinate moiety. 18. The use of a compound according to any one of claims 1 to 7 and 14 for the manufacture of a medicament for the treatment of cell proliferative disease. Priority from 10.29.2003 according to claims 1-9, 11, 14-18 dated October 26, 2004 according to claims 10 and 13.

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